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Human Reproduction, Vol.26, No.5 pp.

1252– 1258, 2011
Advanced Access publication on February 20, 2011 doi:10.1093/humrep/der017

ORIGINAL ARTICLE Reproductive genetics

The p53-HDM2 gene – gene
polymorphism interaction is associated
with the development of missed
abortion
Yan Fang 1, Beihua Kong 1,*, Qifeng Yang 2, Daoxin Ma 3, and Xun Qu 4
1

Department of Obstetrics and Gynecology, Qilu Hospital, Shandong University, 107 Wenhua Xilu, Ji’nan, 250012 Shandong, China
Department of General Surgery, Qilu Hospital, Shandong University, 107 Wenhua Xilu, Ji’nan, 250012 Shandong, China 3Department of
Hematology, Qilu Hospital, Shandong University, 107 Wenhua Xilu, Ji’nan, 250012 Shandong, China 4Department of Basic Medicine,
Qilu Hospital, Shandong University, 107 Wenhua Xilu, Ji’nan, 250012 Shandong, China
2

Submitted on September 3, 2010; resubmitted on December 22, 2010; accepted on January 12, 2011

background: The interaction between p53 and human double minute 2 (HDM2) plays an important role in apoptosis; therefore, functional polymorphisms in these genes might have adverse effects in early pregnancy. In this study, we investigated whether p53 codon 72 and
HDM2 promoter (SNP309) polymorphisms were associated with the development of missed abortion.
methods: Women with missed abortions (n ¼ 60) and healthy controls (n ¼ 64) were included in the study. Genotyping of the p53
codon 72 and HDM2 SNP309 (T . G) polymorphisms was performed by PCR with sequence-specific primers and PCR-restriction fragment
length polymorphism analysis, respectively, using villous samples. The mRNA and protein levels for p53 and HDM2 were measured by realtime PCR and semi-quantitative immunohistochemistry, respectively.
results: For the p53 codon 72 polymorphism, no difference in genotype or allele frequencies was observed in women with missed abortion versus controls. However, for the HDM2 SNP309 (T . G) polymorphism, G/G genotype was associated with a higher risk of missed
abortion compared with the T/T + T/G genotypes (P ¼ 0.043). Women carrying the HDM2 G/G genotype or p53 Pro/Pro genotype had
higher HDM2 mRNA (P ¼ 0.04 and P ¼ 0.013, respectively) and protein (P ¼ 0.001 and P ¼ 0.037, respectively) levels than women
with other HDM2 SNP309 and p53 codon 72 genotypes.
conclusions: The genotypes HDM2 SNP309 G/G and p53 codon 72 Pro/Pro can induce high levels of HDM2, which may be associated with missed abortion.
Key words: missed abortion / p53 / human double minute 2 / polymorphism / pregnancy

Introduction
Missed abortion refers to a pregnancy in which there is a fetal
demise without outside intervention but the uterine activity is
absent to expel the products of conception before 20 weeks of gestation (Griebel et al., 2005). Appropriate apoptosis has been shown
to be critically important for the successful development of normal
pregnancy (Chatzaki et al., 2001; Jerzak et al., 2002; Savion et al.,
2002; Choi et al., 2003). Spontaneous pregnancy disorders are
associated with the excessive apoptosis of trophoblasts (Halperin
et al., 2000).
p53, as an important tumor suppressor gene, is called the ‘gatekeeper
of cellular genome’. p53 expression has been demonstrated in first trimester placenta (Quenby et al., 1998). When a cell is exposed to DNA

damage, p53 can induce cell cycle arrest in G1 phase (DNA presynthetic
phase) and apoptosis. Increased human double minute 2 (HDM2;
human ortholog of murine double minute 2) levels were shown to
lead to attenuation of the p53 DNA damage response, and HDM2 is
a key negative regulator of p53 (Zauberman et al., 1995; Ries et al.,
2000). Enhanced p53 expression and activity subsequently leads to
induction of HDM2, which may act as an oncogene (Fakharzadeh
et at., 1991; Dubs-Poterszman et al., 1995; Jones et al., 1998) or to
inhibit growth (Brown et al., 1998). Specifically, enhanced HDM2
levels have been shown to cause p53 degradation in surviving cells
and, consequently, attenuation of the p53-mediated DNA damage
response (Zauberman et al., 1995; Ries et al., 2000), thereby forming
a negative feedback loop (Michael et al., 2003).

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*Correspondence address. Tel: +86-531-82169008; Fax: +86-531-86927544; E-mail: kongbeihua@yahoo.com.cn

Fetal cardiac activity and gestational age were confirmed by ultrasound.. R: 5′ -CGT GCA AGT CAC AGA CTT-3′ . the patient was considered as heterozygote (Arg/Pro). Based on the fact that the p53-HDM2 interaction plays an important role in apoptosis. Patients This prospective observational study involved 60 pregnant women diagnosed with first-trimester missed abortion. DNA samples were routinely stored at 2208C.. while the Pro-p53 could induce a higher level of G1 arrest than the Arg-p53 (Pim et al. Genotyping of p53 codon 72 and HDM2 SNP309 PCR analysis of p53 codon 72 polymorphism Analysis of p53 genotype at codon 72 was performed as described (Pietrowski et al.. 26 had at least two prior miscarriages. PCR conditions for the Pro allele were as follows: 948C for 5 min.. 2005).25 U Taq Platinum Polymerase (Tiangen). we found that the HDM2 SNP309 G/G genotype was associated with a high risk of missed abortion (Fang et al. Several PCR products were sequenced to further validate the PCR results. 528C for 50 s. immunologic factors (such as positive anticardiolipin antibody and positive antinuclear antibody) and infections. Villous samples (n ¼ 64. Importantly. None of these women had had a successful pregnancy. PCR conditions for the Arg allele were as follows: 948C for 5 min. All the pregnancies terminated at the first trimester . this HDM2 SNP309 G/G polymorphism enhanced promoter recognition by the transcription factor Sp1. 2004). 2003). Recently. Written informed consent was obtained from all participating subjects. 728C for 50 s and a final extension of 728C for 10 min. The gels were stained with ethidium bromide and photographed using an ultraviolet light transilluminator. Dumont et al. the patient was considered as Arg homozygote (Arg/Arg). one per woman) from the missed-abortion group were collected by curettage or manual vacuum aspiration. show a heightened HDM2 expression and significant attenuation of the p53 pathway compared with those carrying the T/T genotype. 2005). (iii) if the sample showed amplification with both two primers. PCR products (10 ml) were subjected to electrophoresis on 3% agarose gels. 10 ng genomic DNA was mixed with 1. 25 mmol/l Tris–HCl (pH 8.10 weeks from the LMP. People’s Republic of China).7). and blood was drawn for testing for chromosomal abnormalities. polymorphism SNP309 (a T to G change at nucleotide 309 in the first intron) was found in the promoter of HDM2.. The control group consisted of 64 women in early pregnancy with a healthy. 5 cycles of 948C for 1 min. In our previous report. 2004. the patient was considered as Pro homozygote (Pro/Pro). Polymorphisms in the p53 and HDM2 genes have therefore been shown to be of functional significance. The DNA content and purity of each sample were analyzed by ultraviolet spectrophotometry (the E260/280 ratio ranging between 1. The p53 codon 72 polymorphism has been proposed as a candidate for increasing the chance of miscarriage in otherwise healthy women (Pietrowski et al. Hong et al. intrauterine gestational sac with the largest diameter exceeding 10 mm but devoid of yolk sac or an empty gestational sac with a confirmed gestational age of no .. HDM2 is vital in the regulation of p53 yet sequence variations in the HDM2 promoter may result in altered expression of the HDM2 protein. Sequence-specific primers (SSPs) for the Pro allele are F: 5′ -GCC AGA GGC TGC TCC CCC-3′ .oxfordjournals. The possible outcomes were: (i) if a PCR product was obtained only with the Pro-specific primers. we hypothesized that the functional polymorphisms in p53 codon 72 and HDM2 SNP309 might be associated with a risk of missed abortion.8). Cells carrying the G/G genotype. and tested this hypothesis in this study . The study design was approved by the Ethical Committee of Shandong University. 2005).1253 p53/HDM2 gene polymorphisms and risk of missed abortion Materials and Methods Diagnosis By ultrasound examination. with these analyses resulting in an unexplained etiology. 1999. R: 5′ -CTG GTG CAG GGG CCA CGC-3′ .. 250 mmol/l each dNTP. Specimens Villous samples (n ¼ 60. Each villous sample was divided into two parts: one part was stored at 280oC before genomic DNA and RNA isolation. all subjects underwent a standard diagnostic work-up to rule out any verifiable cause of missed abortion. the first-trimester missed abortion was defined as an intact gestational sac lacking any fetal cardiac activity [6 weeks after last menstrual period (LMP)]. 648C for 45 s. these forms are ascribed to amino acid replacement at codon 72 of Arg (CGC) by Pro (CCC) in the transactivation domain of the p53 protein. 10 mmol/l KCl. Beijing. randomly selected) was stored in 4% formaldehyde at room temperature overnight for immunohistochemistry analysis. The Pro product was 178 bp and the Arg product was 136 bp. In each 25-ml reaction. 2 mmol/l MgCl2 (Tiangen) and 10 mmol/l of each primer (previously described). At least two forms of wild-type p53 protein exist among human populations.6 and 1..org/ by guest on November 14. Among the 60 women who suffered from missed abortion.6 weeks (Griebel et al. viable intrauterine fetus and no prior miscarriage. 2015 Single nucleotide polymorphisms (SNPs) in the p53 gene might have functional relevance. the other part (only 30 from missed-abortion group and 31 from control group. owing to an enhanced affinity for binding the stimulatory protein (Sp) 1. PCR-restriction fragment length polymorphism analysis of HDM2 SNP309 polymorphism Primers used in this study have been described: 5′ -CGCGGGAGTT CAGGGTAAAG-3′ and 5′ -AGCTGGAGACAAGTCAGGACTTAAC-3′ Downloaded from http://humrep. 728C for 50 s and 35 cycles of 948C for 1 min. DNA preparation DNA was extracted using a DNA isolation kit (Tiangen. 728C for 50 s and a final extension of 728C for 7 min. The arginine and proline isoforms have different biological and biochemical effects: the Arg-p53 could induce apoptosis more efficiently than the Pro-p53 (Thomas et al. 618C for 50 s. including DNA damage (Bond et al. 24 had one prior miscarriage and 10 experienced miscarriage for the first time. 2005). which in turn caused elevated HDM2 expression and attenuation of the p53-mediated apoptotic response to cellular stresses. (ii) if a PCR product was obtained only with the Arg-specific primers.. and a 10 ng DNA aliquot of each sample was used for PCR amplification. one per woman) from the control group were obtained by vacuum aspiration from women undergoing elective abortion at 7 – 10 weeks of gestation for social reasons. 35 cycles of 948C for 1 min. The women were examined using ultrasonography for uterine abnormalities. 2009). Prior to inclusion in the study. Primers of Arg allele are F: 5′ -TCC CCC TTG CCG TCC CAA-3′ .

767. The OD260/280 of the RNA samples ranged between 1.636). Significant difference was defined as P .330. 1.1254 (Ohmiya et al. was carried out using the Roche sequence detection system (Roche. (2001). and reverse: 5′ -ATCTGTTGCAATGTGATGAAG-3′ . there was also no difference between women with missed abortion and controls (P ¼ 0. . The quantity of RNA was assessed spectrophotometrically. 95% CI. 2. Mouse monoclonal antibody for wild-type p53 (Zhongshan. 1 ml RNAse inhibitor. Immunohistochemistry Villous samples (61: 30 from women with missed abortion and 31 from controls) were fixed in 4% formaldehyde overnight at room temperature. Beijing. the patient was considered as T homozygous (T/T). and 708C for 10 min).472–1. Tissue sections were deparaffinized and rehydrated through graded alcohol.5% immunopositive cells as +.054. . diluted 1:50) and HDM2 antibody (Zhongshan. embedded in paraffin wax and four-micron sections were cut (Zhongshan. Chi-squared test was also used to analyze the association of p53 and HDM2 polymorphisms with the immunohistochemistry results for p53 and HDM2. 1 ml. 588C for 10 s and 728C for 10 s and fluorescence reading taken at 728C. People’s Republic of China) was used for the quantification of p53 and HDM2 mRNA expression. People’s Republic of China). The reagents were the same as for PCR-SSP above. The total RNA was reverse transcribed into cDNA using the Rever Tra Ace Kit (Takara) in a volume of 20 ml. After DNA restriction. OR. 189 and 48 bp were observed. Shanghai. Total RNA was extracted from villous samples using the Trizol reagent (Takara. The mRNA levels of p53 and HDM2 in villous samples The relationship of the p53 codon 72 and HDM2 SNP309 polymorphisms to the levels of p53 and HDM2 mRNA were examined in villous samples (n ¼ 124). 1 ml Rever Tra transcriptase.282). 0. The PCR conditions for HDM2 SNP309 were as follows: 948C for 7 min..778. 0.5 mg/ml) and 5 ml ample RNA in a thermal cycler (room temperature 10 min. Cycling parameters were the following: denaturation for one cycle at 958C for 10 s. The gels were stained with ethidium bromide and photographed using an ultraviolet light transilluminator. The odds ratio (OR) was used to measure the association between the allele frequencies and risk of missed abortion. 35 cycles of 948C for 40 s. All the values were presented as mean + SD. The percentages of p53. Shanghai. The sections were incubated with biotinylated goat anti-rabbit immunoglobulin (Ig) G for 20 min at 378C and then processed according to the SP kit (Zhongshan) protocol. After three rinses with phosphate-buffered saline. Endogenous peroxidase activity was quenched by incubation in 3% hydrogen peroxide for 10 min. The primers used for HDM2 were: forward: 5′ -GGACAAGAACTCTCAGAT GAAGATG-3′ . In the negative control for staining. 3′ ) (NEB. The assay for the HDM2 polymorphism utilized the enzyme MSPA1I (5′ . The PCR mixture consisted of 10 mM of each primer. 95% CI. (iii) if three DNA fragments of 237. including 4 ml of 5 × reverse transcriptase buffer. 0. . 45 cycles (temperature transition of 208C/s) of 958C for 0 s. The absence of any positive cells was scored as negative. 728C for 40 s and a final extension of 728C for 7 min. Results The genotypes and allele frequencies of p53 codon 72 polymorphism Arg/Pro (60%) was a dominant genotype.org/ by guest on November 14. Student’s t-test was used to compare the p53 and HDM2 mRNA levels in villous samples. Real-time PCR analysis of p53 and HDM2 mRNA The Quantitative SYBR-Green PCR mix kit (Toyobo. 1 ml Oligo (dT) 18 (0. The mRNA samples were stored at 2808C before analysis. CNGCKG .668.and HDM2-positive cells were estimated by counting the number of immunoreactive cells in five microscopic fields at high-power magnification (×400) in areas with the highest visually determined positive immunoreactivity. . Dalian. Individual p53 and HDM2 measurements was calculated relative to expression of GAPDH using a modification of the method described by Lehmann et al.00. while the frequency of Pro/ Pro genotype was only 20%. diluted1:50) were applied to the sections separately overnight at 48C. . 1. sections were incubated with isotype-matched non-specific antibodies (MsIgG). 2 ml dNTP (10 mmol/l).661–4.092–7. sections were blocked with normal goat serum to suppress non-specific background staining. 608C for 40 s. The possible outcomes were: (i) if only one DNA fragment of 237 bp was observed. The genotypes and allele frequencies of HDM2 SNP309 polymorphism The G/G genotype was more in women with missed abortion (28. the patient was considered as G homozygous (G/G).oxfordjournals. A higher frequency of the G allele was detected in women with missed abortion (48.373.80 and 2.33%) than controls (35. 5 – 20% as ++. OR.773). we used a complete DNA amplification mix in which the target cDNA template was replaced by water. For comparisons of the frequency of Arg and Pro alleles. the patient was considered as heterozygous (T/G). a significant difference was seen between women with missed abortion and controls (P ¼ 0.33%) than controls (12. 10 ml Mix SYBR Green I (Toyobo) and 500 ng cDNA to a final volume of 20 ml.043.05. and reverse: 5′ -CCTCATTCAGCTCTCGGAACATC-3′ .003–2. 2015 RNA extraction and RT reaction Fang et al. with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was an internal reference gene. In the comparison of frequency of T/T + T/G and G/G genotypes. 1. The primers used for p53 were: forward: 5′ -ACTAAGCGAGCACTGCC CAAC-3′ .750. For negative controls.011). People’s Republic of China). Nuclear staining of cells was regarded as a positive result.50%).94%) (P ¼ 0. and melting curve analysis with continuous fluorescence reading. Guangzhou. Relative gene expression quantification for p53 and HDM2. 2006). 95% CI. No significant difference in frequency was observed between the Pro/Pro and the Arg/Arg + Arg/Pro genotypes in women with missed abortion versus controls (P ¼ 0. OR. 0. All P-values are two-tailed and 95% confidence intervals (CIs) were calculated. samples from women with the Pro/Pro genotype and G/G genotype had a higher HDM2 mRNA Downloaded from http://humrep. Statistical analysis Chi-squared test was used to analyze the genotype distribution of p53 codon 72 and HDM2 SNP309 polymorphisms between women with missed abortion and control groups. OR. 95% CI. Several PCR products were sequenced to further validate the PCR results. . (ii) if only two DNA fragments of 189 and 48 bp were observed. 428C for 1 h. 10 ml of digested samples were subjected to electrophoresis on 3% agarose gels. . 1. People’s Republic of China). People’s Republic of China). In the comparison of the p53 codon 72 and HDM2 SNP309 polymorphisms.20% as +++.

In the comparison of the Arg and Pro alleles. all Student’s t-test.001) but there was no difference in protein levels for p53 in comparison of T/T genotype with the T/ G + G/G genotypes (P ¼ 0. including 21cases of T/T genotype.278 and P ¼ 0. 2003). (b) Comparison of mRNA levels for HDM2 between genotypes p53 Pro/Pro and Arg/Arg.org/ by guest on November 14. However. 2) p53 codon 72 genotype and the expression of p53 and HDM2 Sixty one villous samples were randomly selected. The G/G genotype had a higher HDM2 level than T/T + T/G genotypes (P ¼ 0. (2006) used an unselected population and Pietrowski et al. 1b and a). (c) Comparison of mRNA levels for p53 between genotypes HDM2 G/G and T/T (P ¼ 0. the HDM2 SNP309 G/G genotype can induce higher HDM2 mRNA levels than the T/T genotype (P ¼ 0. no significant difference was observed in frequency of the Pro/Pro and the Arg/Arg + Arg/Pro genotypes between women with missed abortion and controls (P ¼ 0. we found that the HDM2 SNP309 G/G genotype was associated with a high risk of missed abortion and concluded that the HDM2 SNP309 G/G genotype may be a genetic risk factor for missed abortion (Fang et al.289). The Pro/Pro genotype had a higher percentage of HDM2-positive cells than the Arg/Arg + Arg/Pro genotypes (P ¼ 0. there was also no difference between women with missed abortion and controls (P ¼ 0. 1d and c). (2005) reported a significant association between the Pro allele and the occurrence of recurrent pregnancy loss. the G/G genotype can induce higher HDM2 mRNA levels than the T/T + T/G genotype (P ¼ 0. The cytotrophoblast cells. gene polymorphisms have been proposed as susceptibility factors that increase the chance of miscarriage in otherwise healthy women (Pietrowski et al.563). syncytiotrophoblast cells and extravillous trophoblast cells showed positive nuclear staining in the villous samples (Fig.04).278).037). into whether the genetic polymorphism of p53 codon 72 and HDM2 SNP309 were associated with the risk of missed abortion. The difference may be related to the different populations used in the studies.. Similarly. As described in our previous report. (a) Comparison of mRNA levels for HDM2 between genotypes HDM2 G/G and T/T (*P ¼ 0.. Missed abortion is a complicated problem. In the present study. Immunohistochemistry Nuclear staining was scored as positive expression for p53 and HDM2. through molecular examination. 35 cases of Arg/Pro genotype and 10 cases of Pro/Pro genotype (Table I). 2015 Sixty one villous samples were randomly selected.04) (Fig. *a – d. 2009).04).013 and P ¼ 0. HDM2 SNP309 genotype and the expression of p53 and HDM2 Discussion Figure 1 p53-HDM2 gene – gene polymorphisms and the mRNA. 24 cases of T/G genotype and 16 cases of G/G genotype (Table II). (d) Comparison of mRNA levels for p53 between genotypes p53 Pro/Pro and Arg/Arg (P ¼ 0. 2005): furthermore.1255 p53/HDM2 gene polymorphisms and risk of missed abortion level than with the Arg/Arg genotype and T/T genotype (P ¼ 0. including 16 cases of Arg/Arg genotype. It has been found that there is significant variation in p53 codon 72 polymorphisms among different ethnicities (Inserra et al. using samples from women with missed abortions and controls. multiplicative interactions between p53 and HDM2 polymorphism genotypes had a higher risk of the occurrence of missed abortion. No significant difference can be seen in the comparison of the Arg/Arg and T/T homozygote with the Pro/Pro and G/G homozygote on p53 mRNA level separately (P ¼ 0. These findings support our prior hypothesis Downloaded from http://humrep. (n ¼ 124).oxfordjournals. Coulam et al. while Coulam et al. in villous samples.037). 2003. no difference can be seen in comparison of Arg/Arg genotype with the Arg/Pro + Pro/Pro genotypes (P ¼ 0.013). Similarly. (2005) included only Caucasian women whose parents were of the same ethnicity. but for p53 positive cells.373). The patient population in our study included only Han people.001). the Pro/Pro genotype can induce higher HDM2 expression than the Arg/Arg + Arg/Pro genotype (P ¼ 0. .. The Pro/Pro p53 codon 72 genotype can induce higher HDM2 mRNA levels than the Arg/Arg genotype (P ¼ 0.013).330). (2006) found no significant differences when comparing genotypic and allelic frequencies of p53 codon 72 among patients with recurrent pregnancy loss and controls.367). Pietrowski et al.367) (Fig.The present study provides new insights. levels for p53 and HDM2. (P ¼ 0.

... T/T 21 14 7 T/G 24 14 10 G/G 16 2 14 0..........289e 0.... 61b 2 or 1 Pa p53 ..............037d 8 8 21 14 6 4 0................. sections were incubated with isotype-matched non-specific antibodies (MsIgG) (C)................... One key regulator may be p53........ Arg/Arg 16 11 5 Arg/Pro 35 17 18 Pro/Pro 10 2 8 0. HDM2 ............000f a All by x 2 test........................... f p53 immunostaining in G/G versus T/T and T/G genotypes................. p53 is also a potential mediator for pregnancy (Sivaraman Downloaded from http://humrep............................... Apoptosis and cell proliferation are coordinately regulated during pregnancy (Pietrowski et al........... HDM2 ........ 2015 Table I p53 codon 72 genotype and the levels of p53 and HDM2 protein....... 11 or 111 2 or 1 P 11 or 111 ............ ................................062c d 0.. Mouse monoclonal antibodies of wild-type p53 (diluted 1:50) and HDM2 (diluted 1:50) were applied to the sections separately (A and B). e p53 immunostaining in T/T versus T/G and G/G genotypes......... f p53 immunostaining in Pro/Pro versus Arg/Arg and Arg/Pro genotypes.. 2005)................1256 Fang et al.563e 1.................. 2001..................... A nuclear yellow– brown stain indicated a positive result............................ d HDM2 immunostaining in G/G versus T/T and T/G genotypes................................. lack of cell contact inhibition and immune privilege (Quenby et al.........155c 0.................... Meanwhile......... 1998)... a potent transcription factor that controls the expression of multiple target genes involved in cell-cycle progression and apoptosis (Sivaraman et al............. 11 or 1 11 2 or 1 P 1 1 or 1 11 ...........oxfordjournals. b Table II HDM2 SNP309 genotype and the levels of p53 and HDM2 protein...................001 10 11 13 11 12 4 0............ high cellular proliferation rate.... as assessed by immunohistochemistry in human villous samples... Thirty randomly selected from women with missed abortions and 31 from control group......... 2005)................... c HDM2 immunostaining in Arg/Arg versus Arg/Pro and Pro/Pro genotypes............. In the negative control for staining......................... d HDM2 immunostaining in Pro/Pro versus Arg/Arg and Arg/Pro genotypes........ c HDM2 immunostaining in T/T versus T/G and G/G genotypes.. The trophoblast cells are similar to malignant cancer cells in terms of their invasiveness....142f a All by x 2 test........ Carvajal et al...... 61b 2 or 1 Pa p53 .. Figure 2 Immunohistochemical staining of p53 and HDM2 (×200) in villous samples of control group................. e p53 immunostaining in Arg/Arg versus Arg/Pro and Pro/Pro genotypes............. b that the genetic polymorphisms of p53/HDM2 and their interactions were associated with missed abortion...................org/ by guest on November 14................ Placental trophoblast invasion and placental reorganization is an ongoing process during pregnancy.... Thirty randomly selected from women with missed abortions and 31 from control group............ as assessed by immunohistochemistry in human villous samples.....

119:591 – 602. The codon 72 polymorphic variants of p53 have markedly different apoptotic potential. Thomas CA. Leu JI. Levy et al. The Arg/Arg genotype seems to induce apoptosis with faster kinetics and to suppress transformation more efficiently than the Pro/Pro genotype (Thomas et al.. A single nucleotide polymorphism in the MDM2 promoter attenuates the p53 tumor suppressor pathway and accelerates tumor formation in humans. and a number of studies have revealed the functional differences of the two polymorphisms (Hong et al. 2000. Kin JW. cell cycle arrest and apoptosis. Acknowledgements The authors thank Qingshen Li and Qiuling Zhu from the Maternal and Child Health hospital of Ji’nan City for help with collecting the blood and villous samples from the pregnancy women. predicting the clinical outcome of advanced non-small cell lung cancer and influencing the clinical outcome of transitional cell carcinoma of the bladder (Horikawa et al. possibly then resulting in the death of the embryo. 2005). The HDM2 G/G carriers might more easily respond to stress.. Kouimtzoglou E. 30872738). The lower level of apoptosis might lead to misguided growth of cells and high expression of HDM2. Murphy M.. Choi BC.. Bargonetti J. we observed that the woman who carry the G/G or Pro/Pro genotype had significantly higher expression levels of HDM2. This study supports our prior hypothesis that polymorphism variations in p53 and HDM2 might have a relationship with missed abortion.. Tovar C. Goldberg et al. In our study. Elevated HDM2 protein binds to p53 to inactivate its transcriptional activity and facilitate the destabilization of p53. Missed abortion may be related to trophoblast cells undergoing severe DNA damage. Arva NC. 2001. Wuerl P et al. Chatzaki E. Michael et al. The investigated polymorphisms of p53 and HDM2 have been shown to be of functional significance.. Margioris AN. Oncogene 1995. thus. Gravanis A. Funding This work was supported partly by National Natural Science Foundation of China (No. Yang H. 1999. Dubs-Poterszman MC. Carvajal D.23:541 – 547. 2002. The major regulator of p53 activity is the HDM2 protein.33:357– 365. The Fas/FasL apoptosis pathway is involved in kappa-opioid-induced apoptosis of human endometrial stromal cells. (2004) showed that after treatment with etoposide to induce DNA damage. 2015 et al. 2006). 2008). our findings support the hypothesis that the polymorphisms of p53 codon 72 and HDM2 SNP309 and their multiplicative gene –gene interaction are associated with an increased risk of missed abortion. Reprod Biomed Online 2006. 2008). 1998. George DL. In our study. leading to the high expression of HDM2. Researchers found that both p53 codon 72 polymorphism and HDM2 SNP309 were associated with the age at onset of oral carcinoma (Yoon et al.. Hu et al. In conclusion. One explanation for the observed increase of missed abortion for p53 Pro and HDM2 G/G carriers might be their higher potential to resist apoptosis. The maintenance of placenta is essential for the exchange between the fetus and its mother and apoptosis and cell proliferation are major processes contributing to a successful pregnancy. with a later onset of response to stress or a lower level of the required physiological apoptosis. MDM2 transformation in the absence of p53 and abrogation of the p107 G1 cell-cycle arrest...1257 p53/HDM2 gene polymorphisms and risk of missed abortion epidemiologic study investigated whether the p53 and HDM2 polymorphisms were associated with a risk of missed abortion. significant death was observed in cells with the HDM2 T/T genotype but not in cells with the HDM2 G/G genotype. 1999. Mayo et al. Taubert H. The high expression of HDM2 can attenuate the apoptosis reaction following DNA damage. 1993). In our study. Zauberman et al. Choi HK. Lutzker SG.This . Heimbrook DC. 2002). which forms a negative feedback loop. Della Pietra ACr. Tocque B. Lee SH. Deb SP..11:2445 – 2449. Vu BT. early development of hepatocellular carcinomar in patients with chronic hepatitis B virus infection (Han et al.. Robins H. Bond GL. 2002).. We suggest that p53 Pro carriers might be at a disadvantage. For the HDM2 polymorphism.oxfordjournals. Mol Reprod Dev 2003. p53 is activated by stressful stimuli.001. enhanced HDM2 levels have been shown to cause p53 degradation in surviving cells and consequently attenuate the p53-mediated DNA damage response (Ries et al. Jeyendran RS. Bond EE. Bond et al... Specifically. Hu W. Carcinogenesis 2002.1:24– 31.04 and P ¼ 0. Ronai Z. the homozygous Pro/Pro genotype was found more often among women with missed abortion compared with controls. EMBO J 1998.. which also feeds back to promote p53 degradation (Alarcon-Vargas et al. Bartel G. 2005. 1995). Wasylyk B.and apoptosis-related genes in chorionic villi derived from recurrent pregnancy loss patients. Nat Genet 2003. respectively). At least two forms of wild-type p53 protein exist among major human population groups. Shandong University for help with statistical analysis. 1998. Activation of p53 by MDM2 antagonists can protect proliferating cells from mitotic inhibitors. Makrigiannakis A. Pietrowski et al. 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