Impact Of MiR-137on Proliferation Of Laryngeal Carcinoma Cell Line Hep-2

Research background：MicroRNAs (miRNAs) are a class of small non-coding RNAs and consist of20-25ribonucleotides. The mature miRNAs with full or partial complementarity tothe target mRNAs direct the cleavage of target mRNAs or act as repressors oftranslation, involved in modulating the development, differentiation, proliferationand apoptosis of cells.Recent studies have shown a distinct connection between miRNAs and thedevelopment of cancer, some of which are specially expressed in glioblastoma. Wehave collected different pathological diagnosis of malignant glioma, normal braintissue as well as glioma cell lines, and then screen the glioma-specific expression ofmiRNAs using miRNAs microarray methods to find specific expression in humangliomas of miRNAs,miR-137is one of them. As an oncomiRs, miR-137, has been reported to bedyregulated in a variety of cancer. In this study, we showed that inprove of miR-137inhibited cell growth in Hep-2cells, accompanying a decreased expression of humancell division cycle42(CDC42蛋白). Taken together, these results suggest thatmodulation of the mechanism responsible for miR-137in Hep-2could be used as acritical therapeutic strategy for Laryngeal intervention and warrants furtherinvestigation.Methods：1. Selected Hep-2cells, which were cultured in10%newborn calf serumDMEM medium, and placed in37℃,5%CO2incubator, taking logarithmic phase for cell experiments. The experiments were divided into antisense miR-137group,scramble group and blank group. The effects of the proliferation, cell cycle,apoptosis and related target genes were evaluated by MTT assay, Flow cytometricmethod.2. Engineered of lentiviral vector based siRNA construct against CDC42anddivided the experiment into CDC42siRNA group, negative control siRNA gruop, andblank group. The effects of the proliferation, cell cycle and apoptosis were evaluatedby MTT assay and Flow cytometric method after the lentiviral vectors infected intoHep-2cells.3.Western blot assay and RT-PCR assay were used to study the expression ofCDC42in Hep-2cells atfer transfecte with antisense miR-137.Results:1.improve of miR-137by antisense oligonucleotide inhibited cell growth,arrested the cell at G0/G1phases, and induced cell apoptosis in Hep-2cell, with adecreased expression of CDC42.2. Knockdown of CDC42expression using lentiviral vector mediated deliveryof CDC42-siRNA triggered cell apoptosis and cell cycle G1/G0arrest in Hep-2cells.improve of miR-137considerably inhibited tumor growth and diminished theexpression of CDC42,Conclusions:1. improve of miR-137by antisense oligonucleotide inhibited cell growthHep-2cells, with a decreased expression of CDC42. rise of miR-137considerablyinhibited tumor growth and diminished the expression of CDC42in xenograftmodel.2. Slow virus mediated CDC42siRNA infection of Hep-2cells, foundCDC42siRNA laryngeal cancer cell proliferation inhibition, prompting block at the G1/G0laryngeal cancer cells, and induced its apoptosis.3. These results indicate for the first time that miR-137inhibits cell growththrough regulating CDC42expression in a dependent pathway in Hep-2cells.