The Relationship Between TRPV6 And PARP-1 In The Process Of Inhibiting SGC-7901 Gastric Cancer Cell Lines' Proliferation With ?-lapachone

Objective:To explore the relationship between TRPV6 and PARP-1 in inhibiting SGC-7901 gastric cancer cell lines proliferation with ?-lapachone.Method:1.The frozen SGC-7901 gastric cancer cell lines were made to come back to life,and then the recovery cell lines were subcultured.After that,MTT assay was used to detect the effective drug concentration of ?-lapachone to SGC-7901 gastric cell lines.2.Three groups of TRPV6-siRNA gene orders were found,including TRPV6-siRNA group 1,TRPV6-siRNA group 2 and TRPV6-siRNA group 3.Each gene order was divided into two concentration,50nM and 100nM.After transfecting SGC-7901 gastric cell lines,Western Blot method was used to select a group of stable transfecting siRNA cell lines which had the best transfection effect compared with the NC-siRNA group.3.SGC-7901 gastric cell lines were divided into two groups,including the group of TRPV6 and the group of PARP-1.The TRPV6 group was further divided into another four groups,including the control group,si-TRPV6 group,?-lapachone group and si-TRPV6 adding ?-lapachone group.The PARP-1 group was divided into three groups,including the control group,?-lapachone group and si-TRPV6 adding ?-lapachone group.Western Blot method was used to test TRPV6 and PARP-1 protein's expression of each group.4.The selected siRNA was used to transfect SGC-7901 gastric cell lines again and then a control group was set up.The cells were treated with 1,2,4,8?mol/L?-lapachone.MTT colorimetric analysis was used to test the cells' viability of each group.Results:1.According to the MTT test,the effective concentration of ?-lapachone was 4.0?mol/L when inhibiting SGC-7901 gastric cell lines took up 50%.2.The result of the Western Blot experiments showed that,compared with NC-siRNA,the expression of TRPV6 protein in 50nM TRPV6-siRNA 1 to 3 transfection groups and in 100nM TRPV6-siRNA 1 to 3 transfection groups decreased.Among them,the expression of TRPV6 protein in 100nM TRPV6-siRNA transfection group 2 decreased most obviously(P<0.05).Therefore,100nM TRPV6-siRNA transfection group 2 was chosen to silence the SGC-7901 gastric cancer cell lines.3.Western Blot test showed that,compared with the cell lines which are not silenced,the expression of TRPV6 protein of the silenced gastric cell lines decreased,indicating that this si-RNA order is effective.Meanwhile,PARP-1 in TRPV6-siRNA transfection group was reduced too(P<0.05).4.MTT test showed that,compared with the control group,the cell survival rate of the TRPV6-siRNA transfection group was increased significantly,with regardless of which concentration of P-lapachone solution.5.Pearson linear correlation analysis showed a significant positive correlation between the expression of TRPV6 protein and PARP-1 protein.Conclusion:1.After inhibiting the expression of TRPV6 protein,the inhibition rate of SGC-7901 gastric cell lines are down-regulated.2.In the process of inhibiting SGC-7901 gastric cancer cells with ?-lapachone,TRPV6 and PARP-1 are mutually coupled,and jointly promote the death of the gastric cancer cell.