Effects Of Silencing PTHrP Gene Expression On Invasion Of SPC-A-1BM Cells By RNAi

Objective:In order to explore parathyroid hormone-related protein(PTHr P)gene silencing impact on the invasive ability of human lung SPC-A-1BM cells and receptor activator of nuclear factor-κB ligand(RANKL)gene expression.Methods : The experiment were classified into five groups : 1-3 groups are the experimental group,si RNA inhibited PTHr P specifically si RNA-PTHr P interference group,design 3 different sequence si RNA;negative control group,si RNA-NC interference;blank control group,no si RNA interference group.SPC-A-1BM cells were transfected by si RNA with nucleic acid transfection reagent liposomes Lipofectamine TM2000(Lipo2000).The invasive ability of SPC-A-1BM cells was detected by the transwell assay after transfection.The expression of PTHr P m RNA before and after transfection were measured by Real-time polymerase chain reaction(Real-time PCR).The expression of PTHr P and RANKL protein before and after transfection were measured by Western-Blot.Results:After 12 hours,the Transwell test showed that the cells of each group through the Transwell artificial membrane and attached to the bottom of the upper chamber,counted and analyzed statistically.The results showed that compared to PTHr P specific transfection group(group1,2,3)and control group(group 4、5),the cell number of passes through the artificial membrane were decreased,the inhibition rates were 63.8%,60.3%,66%.By the single factor analysis of variance,it showed that the experiment group1,2,3 and control group 4,5 was statistically significant(P < 0.01),between the group 4and 5 was no significant difference(P > 0.05);Real-time PCR showed that experimental group1,2,3 of PTHr P gene the inhibition rate was significantly inhibited,respectively72%,75%,73%,P < 0.01,while the control group,group 4 and 5 comparison,there were no significant differences in the expression of the PTHr P gene;Western-Blot showed that the PTHr P protein,the experimental group 1,2,3 gray-scale value of straps was no statistically significant(P > 0.05),between the group 4 and 5 was no significant difference(P > 0.05).No matter in the group 1,2 or 3,the gray values of straps were statistically significant compared with groups 4,5(P < 0.01).The similar to the RANKL protein,the experimental group 1,2,3 gray-scale value of straps was no statistically significant(P >0.05),between the group 4 and 5 was no significant difference(P > 0.05).No matter in the group 1,2 or 3,the gray values of straps were statistically significant compared with groups 4,5(P < 0.01).Connclusion : The experimental findings indicated that PTHr P gene silencing can inhibit the invasion ability of SPC-A-1BM cells and down-regulate the expression of RANKL significantly;A new target for the treatment of lung carcinoma is supplied bythis work.