| Background and objectiveThe incidence of cervical carcinoma accounted for the first in gynecologicalmalignancies.The pathogenesis of cervical carcinoma has not been fullyelucidated.but,it has relationship with a long term infection with human papillomavirus(Human papillomavirus,HPV).But,not all women infected with HPV developcancer.The occurrence and development of cervical cancer is an extremely complexprocess,involving abnormal structure and expression of gene in cells biologicalprocesses.People studied the mechanism of cervical cancer development anddiscovered that microRNA(miRNA,mRNA) deregulation played a critical role intumorigenesis.MicroRNAs are small single-stranded RNA molecules of about20to25nucleotides that do not code a protein and evolute conserved with regulatoryfunctions.The mature miRNA involved in cell proliferation,differentiation,development and apoptosis process.The role of these small RNA is similar to theoncogenes or tumor suppressor genes.For example,the miR-15a/16-1cluster isdownregulated or deleted in leukemic cells of patients with chronic lymphocyticleukemia(CLL);miRNA let-7is low expressed in breast cancer and lung cancer.The expression level of miR-17is upregulated in lung cancer.Wang and others found that,the epression levels of miR-126,miR-143,miR-154were significantly down regulatedin cervical cancer, while the expression levels of miR-15b,miR-16,miR-146a,miR-155were upregulated in cervical cancer.It was found thatmiRNA-143was frequently deregulated in colorectal cancer and increased theexpression level of miR-143in colon cancer cells can inhibit tumor cell proliferationand promote apoptosis.However,the research about the role of miR-143in cervicalcancer has just started.Our previous studies have shown that miR-143is downregulated in the tissue of carcinoma of cervix,the expression level of miR-143hasrelationship with histological grade and lymph node metastasis in cervical cancer.Inthis study,we examined the expression profiles of miR-143before and aftertransfection in human cervical cancer HeLa cell by quantitative RT-PCR,andanalyzed cell growth in vitro by MTT assay.We aimed to explore if theoverexpression of miR-143have an impact on the proliferation of HeLa cell andinvestigated the role miR-143palyed in the pathogenesis of cervical cancer.Ourstudies provide a theoretical basis for the possible clinical application of thesemiRNAs,including genetic diagnosis and molecular biology treatment of cervicalcancer.Materials and methods1. GroupingAccording to the pre-experimental results,we transfected two concentrations ofmiR-143mimics into HeLa cell.Therefore,two experimental groups were set up.Inexperimental group A and experimental group B,HeLa cell was transfected withmiR-143mimics(at a final concentration of10nmol/L and20nmol/L).Three controlgroups were set up.Transfection reagent control group contained transfectionreagent,culture medium,and HeLa cell;blank control group contained culture mediumand HeLa cell;In negative control group,HeLa cell was transfeced with a syntheticmiR-143negative control(at a final concentration of10nmol/L).2. Experimental methodsThe cervical adenocarcinoma cell line HeLa was adopted.According to the abovegrouping,synthetic miR-143(miR-143mimics)was transfected into HeLa cell by transient transfection method to upregulate the expression level of miR-143in HeLacell.The expression changes of miR-143in the HeLa cell was measured by afluorescent quantitative PCR assay in each group.Methyl thiazolyltetrazolium(MTT)was used to analyze the proliferation of the HeLa cell aftertransfection.We aimed to explore the effect of miR-143on the proliferation ofcervical carcinoma HeLa cell.Date were processed using SPSS17.0statisticalsoftware,the numerical variable data were presented as mean±standard deviation (x±s). Student T-test was used to compare the difference between twogroups.Differences of cell proliferation rate among multiple groups were analyzedusing One way analysis of variance for repeated measurement data.Differencesbetween the two groups were compared using LSD method. Inspection level is α=0.05, P <0.05was considered as statistically significant.Results1. Compared with the blank control group,the miR-143relative expression level ofexperimental group A(transfected with10nM miR-143mimics)was7.38and themiR-143relative expression level of experimental group B(transfected with20nMmiR-143mimics)was13.71,the difference between the two groups was statisticallysignificant(P<0.05);compared with transfection reagent control group,the miR-143relative expression level of experimental group A(10nM)was4.23and the miR-143relative expression level of experimental group B was7.86,the difference between thetwo groups was statistically significant (P<0.05).Compared with the experimentalgroup A,the miR-143relative expression level of group B was significantlyincreased,and the difference between the two groups was statistically significant(P<0.05).2. Compared with the three control groups,the proliferation rate of HeLa cell wasdecreased in experimental group A(transfected with10nM miR-143mimics)andexperimental group B(transfected with20nM miR-143mimics),and the differencewas statistically significant(P<0.05);Cell proliferation rate began to decline from the24h post-transfection,after48h,HeLa cell proliferation rate declinedsignificantly.Compared with experimental group A,the HeLa cell proliferation ratedeclined more significantly in experimental group B,the difference between the two groups was statistically significant(P<0.05).Conclusions1. Transfected HeLa cell with miR-143mimics can effectively upregulate theexpression level of miR-143in cervical carcinoma HeLa cell.After transfected withmiR-143mimics,the expression level of miR-143in cervical carcinoma HeLa cellwas significantly increased in experimental A group and experimental group B.Thiseffect was positively related to the transfection concentration.2. After upregulated the expression level of miR-143in cervical carcinoma HeLacell,the proliferation rate of HeLa cell was significantly declined,it has a dose-effectrelationship between the declined degree and transfection concentration.3. The expression level of miR-143in cervical cancer HeLa cell was significantlydecreased, miR-143may play a role similar to tumor suppressor genes in HeLa cell. |
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