Study Of Apopotin-constructed Oncolytic Adenovirus ATV On Human Cervical Carcinoma HeLa Cells

Purpose: With human cervical cancer HeLa cells as a starting point,to investigate the effects on autophagy and apoptosis of human cervical cancer HeLa cells treated with ATV of tumor specific cytotoxicity and replication competent,for the clinical treatment of cancer to provide new treatment ideas and strategies.Methods: At various time points after infection,WST-1 assay measured effect of ATV infection on proliferation of HeLa cells;Annexin V and Hoechst staining were used to detect the ability of oncolytic adenovirus ATV to induce HeLa cell apoptosis;Effect that detection of the cell cycle and cell apoptosis of ATV infected HeLa cells by flow cytometry;Expression levels of autophagy-related protein LC3 that oncolytic adenovirus ATV induced was detected,Beclin-1 by RT-qPCR,and Western Blotting;MDC staining and IFA detected oncolytic adenovirus ATV induced HeLa cell autophagy levels ability;LC3 and Beclin-1 were knocked down by RNAi to detect regulating autophagy-related molecule in ATV on HeLa cells.Results: In vitro,ATV infection inhibited the proliferation of HeLa cells,which is a typical pathway of mitochondrial apoptosis.;PI/RNase showed ATV can block the S phase of HeLa cell cycle,delay the entire cell cycle and inhibit HeLa cell proliferation;ATV can induce and promote autophagy in HeLa cells and stronger than MOCK,inhibition of autophagy for 24 hours,plays an important role in cell apoptosis,therefore,suspected death form from autophagy to apoptosis;through RT-qPCR,Western Blotting,MDC staining and IFA detection.After siRNA(knocked down Beclin-1)transfected HeLa cells,LC3 autophagy pathway decreased and inhibited autophagy,resulting in autophagosomes reduction,ATV infected(knocked down Beclin-1 and LC3)6 h on HeLa cells,ATG5,ATG12 and ULK1-related autophagy pathway levels were increased,revealed ATV-induced HeLa cell autophagy may not completely dependent on LC3 and Beclin-1 pathway.Conclusion: Apoptin-constructed oncolytic adenovirus ATV can promote the autophagy and apoptosis of human cervical cancer HeLa cells and then specifically kill tumor cells.