Effect Of Lentivirus-mediated PDCD11 Gene Silencing On Proliferation,Migration And Apoptosis Of Colon Cancer Cells

Objective: To investigate the effect of PDCD11 gene on proliferation,migration and apoptosis of colon cancer cells by silencing PDCD11 gene expression with lentivirus as a vector using RNA interference technology,and to add new targets for the treatment of colon cancer.Methods: We cultured RKO and SW480 colon cancer cells in vitro and detected PDCD11 gene expression abundance ΔCT in both colon cancer cells using q RT-PCR,and screened the colon cancer cells with the highest PDCD11 gene expression abundance ΔCT for PDCD11 gene silencing assay.Against PDCD11 gene,2 structurally different RNA interference targets were designed and sh PDCD11(PSC47374)and sh PDCD11(PSC47375)RNA interference lentiviral vectors and 1 negative control RNA interference lentiviral vector sh Ctrl were constructed,both lentiviral vectors carried green fluorescent protein gene(GFP)and validated for packaging.The infected colon cancer cell lines were divided into experimental groups sh PDCD11(PSC47374)and sh PDCD11(PSC47375)and control group sh Ctrl.The green fluorescence of colon cancer cells in each group was observed by fluorescence microscopy.After 80% of the cells were infected,the PDCD11 m RNA expression of colon cancer cells was detected by q RT-PCR,and the experimental groups with the highest silencing efficiency were selected for subsequent cell experiments.The Celigo cell analyzer was used to test the effect of silencing PDCD11 gene expression on the proliferation of colon cancer cells for 5 consecutive days in both groups.Transwell migration assay was used to observe the two groups of cells to examine the effect of silencing PDCD11 gene expression on the change of migration ability of colon cancer.The effect of silencing PDCD11 gene expression on apoptosis of colon cancer was examined by Annexin V-APC single-staining assay.Results: The PDCD11 gene expression abundance of two types of colon cancer cells was detected by q RT-PCR,and the results showed that the ΔCt value of RKO cells was 7.74±0.087 and that of SW480 cells was 8.25±0.142.The expression abundance of RKO cells was the highest,and the RKO cell line was selected for the gene silencing experiment.Two PDCD11-interfering lentiviral vectors were constructed and successfully infected with colon cancer RKO cells.The PDCD11 m RNA silencing efficiency of each group of colon cancer cells was detected by q RT-PCR.Compared with the control group,the gene knockdown efficiency of sh PDCD11(PSC47374)in the experimental group reached 71.1%,and that of sh PDCD11(PSC47375)in the experimental group reached 80.4%(P<0.01),and the experimental group was selected Celigo cell counting assay showed that the proliferation rate of experimental group sh PDCD11(PSC47375)cells was significantly inhibited compared to control group sh Ctrl(P<0.01).transwell migration assay showed that compared to control group sh Ctrl,the proliferation rate of experimental group sh PDCD11(PSC47375)cells was significantly inhibited compared to control group sh Ctrl(P<0.01).The Annexin V-APC single-staining assay showed that the number of apoptotic cells in sh PDCD11(PSC47375)cells was increased compared with the control sh Ctrl group(P<0.01).Conclusion: Silencing of PDCD11 gene expression can reduce the proliferation and migration ability of colon cancer RKO cells,and promote the apoptosis of colon cancer RKO cells.