Effects Of GRK5 On Cell Biology Of Non-small Cell Lung Cancer

Objective:Lung cancer is the leading cause of cancer-related deaths worldwide,and non-small-cell Lung cancer(NSCLC),which accounts for about 80%of all cases,is the main subtype of Lung cancer.As lung cancer is prone to recurrence and metastasis,chemoradiotherapy and molecular targeted therapy are the main therapeutic options for advanced non-small cell lung cancer currently.Among them,molecular targeted therapy has the advantages of small toxic and side effects,better curative effect,and significantly improved the quality of life of patients in clinical treatment.Although targeted drugs have prominent advantages in the treatment of tumors,molecular targeted therapy is only applicable to patients with specific gene mutations and is prone to drug resistance,while it is still do not have better treatment for the population without gene mutations.Therefore,the exploration of new target drugs has become a research hotspot in the treatment of lung cancer.Immunohistochemical analysis of non-small cell lung cancer tissue samples in the early stage showed that GRK5(G-protein coupled receptor 5)was highly expressed in lung cancer tissues compared with paracancer tissues.Consistent with the immunohistochemical results,qrt-pcr was used to compare the transcriptional expression levels of GRK5 in lung cancer with adjacent tissues,and also compare beas-2b with other normal lung epithelial cell lines.It was found that GRK5 was indeed highly expressed in lung cancer tissues and cell lines.Therefore,GRK5 plays an important role in NSCLC.GRK5 has been shown to play a key role in the development and progression of multiple pathological conditions,including cancer.In this study,the expression of GRK5 in non-small cell lung cancer cells was detected to explore its biological significance and correlation.Meanwhile,we observed the biological effects of GRK5 on the proliferation,migration and invasion of H1299 and SPC-A1 cells of NSCLC by inhibiting the expression of GRK5 in these two cell lines.To explore the mechanism of its molecular expression,so as to take GRK5 as a new target for the diagnosis,treatment and prognosis of NSCLC in the future.Methods:1.Take GRK5 as the target to synthesize segments of inhibitor and transfected into H1299 and SPC-A1 cells by liposome method,at the same time,set a negative control group.2.By the mean of fluorescence and RT-PCR screening of stable transfection cell line.And by using Western Blot to detect the effect of inhibiting GRK5 expression on GRK5 protein expression in H1299 and SPC-A1 cells.3.To detect proliferation ability in H1299 and SPC-A1 cells with GRK5 inhibitors transfected compared with negative control by CCK8.4.To identify the effect of GRK5 on the migration and invasion in H1299 and SPC-A1 cells by Transwell assay.5.With the method of Cell scratch test identify the migration of GRK5 inhibitors group and negative control group.Results:1.The result of qRT-PCR shows that GRK5 siRNA can effectively decrease the expression of GRK5 in H1299 and SPC-A1 cells(p<0.05).2.Western Blot results showed that GRK5 protein expression was decreased after transfection with GRK5 siRNA(p<0.05).3.CCK8 assay showed that the proliferation of H1299 spc-al lung cancer cells was significantly inhibited after transfection with GRK5 siRNA(p<0.05).4.The cell scratch experiment showed that the migration rate of H1299 spc-al lung cancer cells transfected with GRK5 siRNA was significantly slowed down(p<0.05).5.Cell migration and invasion experiments showed that the migration rate of H1299 spc-al lung cancer cells transfected with GRK5 siRNA was significantly slowed down(p<0.05).Conclusions:1.GRK5 is highly expressed in NSCLC cells.2.Interfering of GRK5 expression can significantly inhibit the proliferation of lung cancer cell lines H1299 and SPC-A1,reduce their migration rate,and promote the apoptosis of lung cancer cells H1299 and SPC-A1.3.In this experiment,we discussed the correlation between GRK5 and the cellular biological function of NSCLC which providing some ideas for new therapeutic targets for NSCLC.