Construction Of Recombinant Lentiviral Vector Of Microrna-29b And Its Inhibit Of Gastric Cancer Cell Migration And Proliferation

Gastric cancer is one of the most common malignant tumor of the digestive system, which originated from gastric mucosal epithelial cells. The incidence of gastric cancer in our country is high,and is ranking first in all types of tumors.Gastric cancer can occur at any age, but the overall trend is the growth with age.The study found that the causes and genetic factors are disease, stomach disease, dietary habits and so on. In recent years found that the incidence of gastric cancer development because of a non-coding RNA, that is microRNAsMicroRNAs (miRNAs) are a class of new discoveries in recent years, which are about 22nt and non-coding RNA molecules, can mediate gene expression through a combination of mRNA's 3'UTR in multicellular organisms to regulate post-transcriptional level. With in-depth study of miRNAs on the function and the number of growing, it has been proved that miRNA play an important role to regulate in an organism's embryo formation, growth and development, cell proliferation, cell differentiation, cell apoptosis, fat metabolism and the development of human disease. The disturbances of microRNAs expression will lead to target genes up or down, however, many of these target genes is correlated with tumors. Thus, miRNAs play an important role in tumor development and progression as the oncogenes or anti-oncogene.In this study, the pMIR-miR-29a/b/c lentivirus vector was generated by PCR amplification of miR-29a/bc precursor sequence. 293TN cells were transfected with pMIR-miR-29a/b/c,Packaging Plasmid(s) and pVSV-G plasmids to package the recombinant lentivirus. The expression vector of mir-29a/b/c Restructuring slow virus have been successfully achieved by tested.HGC-27 cells were infected with recombinant lentivirus and the expression level of miR-29a/b/c was measured by real-time PCR.The expression of target gene MCL-1 was tested by western blot .The results show miR-29a/b/c is overexpressen HGC-27 cells and its target gene expression of MCL-1 was inhibited significantly.T Finally, scratch test and cell proliferation were used to observe the change in cell migration and proliferation after miR-29a/b/c was over-expressed in gastric cancer cells. The results show, The miR-29a/b/c that is overexpressed can inhibit the ability of migration and proliferation of HGC- 27. ( P<0.01)。...