Effect and mechanism of ITGA3 on paclitaxel resistance in non-small-cell lung cancer
Objective To study the effect of integrin subunit α3(ITGA3)on paclitaxel resistance in non-small-cell lung cancer(NSCLC)and its mechanism.Methods Tumor tissues from 50 NSCLC patients treated with paclitaxel com-bined with cisplatin chemotherapy,A549/Tax(paclitaxel-resistant human lung adenocarcinoma)cells,human A549 cell line and human embryonic lung fibroblasts(MRC-5)were selected.RT-qPCR was used to detect the expression levels of ITGA3 mRNA in tissues and cells.A549/Tax cells were used as the research objects and were transfected with pcDNA3.1-ITGA3 plasmid(oe-ITGA3)and pcDNA3.1 empty vector(oe-NC),silencing ITGA3 small RNA(sh-ITGA3)and nega-tive control(sh-NC),respectively,which were recorded as the oe-ITGA3 group,oe-NC group,sh-ITGA3 group and sh-NC group,respectively.Untreated A549/Tax cells were used as the control group.CCK-8 was used to detect the OD450 val-ues at 24 and 48 h.Western blotting was used to detect the expression of EMT-related proteins[vimentin,E-cadherin,snail,]and TGF-β,and immune escape protein[programmed death ligand-1(PD-L1)];RT-qPCR was used to detect the expression of ITGA3 mRNA in A549/Tax cells;Transwell assay was used to detect the migration and invasion abilities of A549/Tax cells.Results After treatment,the expression of ITGA3 mRNA in the tumor tissues was higher than that be-fore treatment(P<0.05).The expression levels of ITGA3 mRNA in the A549 cells and A549/Tax cells were higher than that in the MRC-5 cells(all P<0.05).After treatment,Vimentin,snail,TGF-β and PD-L1 in the tumor tissues of NSCLC patients increased in comparison with those before treatment,while E-cadherin decreased(all P<0.05).Compared with the control group and the oe-NC group,the OD450 values at 24 and 48 h,the number of cell migration and invasion,the IT-GA3 mRNA expression,the expression levels of Vimentin,snail,TGF-β and PD-L1 increased,and the expression of E-cadherin decreased in the oe-ITGA3 group(all P<0.05).Compared with the control group and the sh-NC group,the OD450 values at 24 and 48 h,the number of cell migration and invasion,the ITGA3 mRNA expression,and the expression levels of Vimentin,snail,TGF-β and PD-L1 decreased,and the expression of E-cadherin increased in the sh-ITGA3 group(all P<0.05).Conclusion Down-regulating ITGA3 expression can inhibit the drug resistance of NSCLC cells to paclitaxel,and its mechanism may be related to the inhibition of TGF-β expression.
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