〔Abstract〕 Objective To investigate the effect of reducing OPTN gene expression on the sensitivity of cells to anticancer drugs by establishing the human SKOV3 cell line stably knocking down OPTN protein with RNA interference technique and human SKOV3 cell line stably knockout OPTN protein with CRISPR/Cas9 gene editing technique. Methods The sequences of shRNA and sgRNA were designed for OPTN gene and ligated with lentiviral vector plasmid pGPU6_GFP_Neo and knockout plasmid PX459 vector to produce recombinant vector. The recombinant plasmid vector and packaging plasmids(psPAX2 and pMD2.G) were co-transfected into 293 T cells for virus packaging. The supernatant of virus was collected to infect SKOV3 cells. SKOV3 cell line with stable knockdown and stable knockout of OPTN gene was obtained.RT-PCR was used to detect the changes of OPTN gene expression in knockdown group. PCR was used to detect the changes of genomic DNA in the knockout group. The expression of OPTN protein was detected by Western blot. The effect of DDP on the proliferation of ovarian cancer cells was detected by MTT. The expression of mRNA of caspase3, Bax and Bcl-2 was detected by qRT-PCR. The expression of caspase3, Bax and Bcl-2 proteins was detected by Western blot. Results Lentiviral vector and gene knockout vector were constructed successfully. In SKOV3 cells,the expression of OPTN protein in SKOV3 cells decreased(knockout cell line) or knocked out completely(knockout cell line).Ovarian cancer cells(SKOV3) with OPTN gene knockdown and knockout were less sensitive to DDP. Compared with the control group, the inhibition rate of DDP on cells was lower(P<0.05,P<0.01), and the effect of gene knockout cells was better. The expression of apoptosis-related genes Caspase3 and Bax mRNA and protein decreased, while the expression of Bcl-2 increased in SKOV3 cells with stable knockdown and knockout of OPTN protein by qRT-PCR and Western blot(P<0.05,P<0.01). Conclusion Reducing or removing the expression of OPTN gene in ovarian cancer can reduce the sensitivity of ovarian cancer cells to the anticancer drug DDP, which may be achieved by affecting apoptosis-related pathways.