| Objective:Endometrial carcinoma(EC) is a kind of malignancies whichderives from endometria. It is one of the three most common of gynecologicmalignant tumors,accounting for20%-30%of malignant tumors of female’sreproductive system.In recent years,the incidence of endometrial carcinoma isrising continuously and showing a younger and younger trend.Althoughendometrial carcinoma has been carried out a great deal of research for mangyears,but its exact etiology is still not clear at present.Therefore,to explore thepathogenesis of endometrial carcinoma and to conquer this disease are theurgent problem to be solved.MicroRNA (abbreviated to miRNA,miR)is akind of non-coding small molecule RNA which is composed of about19-25nucleotide and through base pairing with the specific target mRNA3＇UTRor coding regions to inhibit the translation of target mRNA or degradatemRNA,in ultimately to inhibit the expression of gene.As early as1993,Leeect found the first miRNA——lin-4in the body of caenorhabditis elegans.Thesecond miRNA——let-7was discovered in2000.Since then, the upsurge ofmiRNA has been set off.The study found that the miRNAs play an importantrole in cell growth, differentiation,proliferation,apoptosis and developmentprocess of tumor.The different miRNA has different regulation function to cellgrowth,while the regulation function of the same miRNA to different cell lineis also different.MiRNA can regulate many oncogenes and tumor suppressorgenes.And abnormal expression of miRNA is up or down in thetumor.Recently,as a member of numerous miRNA genome,miR-200family aretaken seriously more and more in oncology.MiR-200family mainly includefive members.They are miR-200a,miR-200b, miR-200c, miR-141andmiR-429.According to the gene location of the coding miR-200family,miR-200family can be divided into two groups which are miR-200a/miR-200b/miR-429and miR-200c/miR-141.In humans,the codinggenes of miR-200a/miR-200b/miR-429are located on chromosome1P36.33,and the coding genes of miR-200c/miR-141are located onchromosome12p13.31.In the process of researding miRNA,wu etc detected10groups of endometrial tissues by using gene chip.Compared with thecontrol group,in endometrial carcinoma tissues17miRNAs were up-regulatedwhile6miRNAs were down-regulated including of miR-429.Liang Jing etcresearched miRNA differential expression profile of endometrialadenocarcinoma tissue by using gene chip and also found that miR-429wereobviously up-regulated.Recently foreign others detect that miR-200family arehigh expression in endometrial carcinoma tissue.After the application ofanti-miR-200family in endimetrial cancer cells,they found that an-ti-miR-429significantly inhibited the growth of Ishikawa cells. The study also foundthat miRNA is closely associated with the P53and P53forms a complexregulatory network in the many kinds of tumors process.P53is the mostin-depth one of the tumor suppressor genes which are researched by tumorrealm at the present.It locates in the short arm of chromosome17,17p13.1.Thegene has the funtion which can adjust transcription,inhibit cell growth andinduce apoptosis.Its mutation and missing cause many tumors.The studyshows that P53is related with clinical stage,pathological type,depth ofmyometrial invasion and lymph node metastasis of the endometrialcarcinoma.And there is reported in the literature that P53is consistent with thelevel of postoperative serum CA125,both union of them can be used as one ofindicators which can prognosis and guide clinical treatment.The intention ofthe experiment is trying to suppress the high expression of miR-429throughtransfecting miR-429antisense oligonucleotide(As-miR-429) to the Ishikawacell of the endometrial carcinoma,then to observe the situation of proliferationand apoptosis,to study the effect of miR-429and P53expression,to explorethe mechanism of miR-429in the development process of endometrialcarcinoma,so as to provide a new theoretical basis for clinical diagnosis andtreatment of endometrial carcinoma. Method:1.Culturing Ishikawa cells with RPWI-1640with containing10%fetal bovine serum at37℃,5%CO2,cell incubator culture.2.For the cellsfusion of90%when transfection,the cultured cells are divided into fourgroups:blank control group,negative control group,AS-miR-429goup andtransfection reagent group.3.The cellular growth activity was assayed byCCK-8after transfection.4.The apoptosis situation was tested byAnnexin-V-FITC/PI.5.The miR-429and P53expression was detected byreal-time PCR.6.The protein expression of P53was measured by westernblot.7.All data were statistically analyzed usig SPSS13.0statistical software.Results: Compared with blank control group and transfection reagentgroup, the cell growth was significantly inhibited in the experimental groupand negative control group.Cell proliferative activity of the negative controlgroup is less than the experimental group. The experimental group wassignificantly increased apoptosis. Apoptosis rate (28.45±2.62)of theexperimental group is higher than that of the control group (3.34±1.02) andthe transfection reagent group (4.19±1.13), lower than the negative controlgroup (41.23±3.62).There were Significant difference between theexperimental group with the other groups （P<0.05）. Real-timequantitative test results showed that the expression of miR-429in theexperimental group was significantly decreased, the expressionlevel of p53mRNA is also decreased. In the negative controlgroup, the expression levels of miR-429and P53was declined. Western blotanalysis results showed that the expression of P53are increased inthe experimental group. Compared with the control group and transfectionreagent group, there was not significant in difference (P>0.05). Inthe negative control group, the protein expression of P53is downregulated.Conclusion: It is confirmed that miR-429is expressed in endometrialcancer Ishikawa cells. AS-miRNA-429can inhibits the expression ofP53mRNA and the expression of cell proliferation,which also caninduce cell apoptosis. But the experimental results have no specificity.Therole of miR-429in the development of endometrial carcinoma and itscorrelation with P53expression remain to be further discussed. |
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