抑制盘状结构域受体1蛋白表达对前列腺癌LNCaP细胞恶性生物学行为的影响

Effect of inhibiting discoid domain receptor 1 protein expression on the malignant biological behavior of prostate cancer LNCaP cells

目的观察短发夹RNA(shRNA)靶向抑制盘状结构域受体1(DDR1)蛋白表达对前列腺癌LNCaP细胞生物学行为的影响。方法将DDR1-shRNA(实验组)与正常对照shRNA(对照组)转染LNCaP细胞,采用Western blot法检测DDR1蛋白的表达,Transwell实验检测细胞迁移能力,CCK-8法检测细胞增殖能力,Western blot法检测雄激素受体(AR)、B细胞淋巴瘤/白血病-xL(Bcl-xL)、增殖细胞核抗原(PCNA)的表达。结果实验组LNCaP细胞中DDR1蛋白表达水平低于对照组(t=16.293,P<0.001)。抑制DDR1的表达可减弱细胞的迁移能力(t=23.142,P<0.001);降低LNCaP细胞24、48、72和96 h的增殖能力(t=9.363,P<0.001;t=11.261,P<0.001;t=13.562,P<0.001;t=14.332,P<0.001);降低AR、Bcl-xL、PCNA蛋白的表达水平(t=19.131,P<0.001;t=23.192,P<0.001;t=13.235,P<0.001)。结论抑制DDR1蛋白可能通过下调LNCaP细胞中的AR表达,抑制其迁移、增殖能力,作用机制可能与Bcl-xL和PCNA表达降低有关。

Objective To observe the effect of short hairpin RNA(shRNA)targeted inhibition of discoid domain receptor 1(DDR1)protein expression on the biological behavior of prostate cancer LNCaP cells.Methods DDR1 shRNA(the experimental group)and normal control shRNA(the control group)were transfected into LNCaP cells.Western blot was used to detect the expression of DDR1 protein,Transwell assay was used to detect cell migration ability,CCK-8 assay was used to detect cell proliferation ability,and Western blot was used to detect the expression of androgen receptor(AR),B-cell lymphoma/leukemia-xL(Bcl-xL)and proliferating cell nuclear antigen(PCNA).Results Compared with the experimental group,the relative expression level of DDR1 protein of LNCaP cells in the control group was reduced(t=16.293,P<0.001).Inhibiting the expression of DDR1 could weaken the migration ability of LNCaP cells(t=23.142,P<0.001);inhibition of DDR1 expression reduced the proliferation ability of LNCaP cells at 24,48,72 and 96 hours(t=9.363,P<0.001;t=11.261,P<0.001;t=13.562,P<0.001;t=14.332,P<0.001);inhibiting the expression of DDR1 could reduce the expression levels of AR,Bcl-xL and PCNA proteins(t=19.131,P<0.001;t=23.192,P<0.001;t=13.235,P<0.001).Conclusion Inhibiting DDR1 protein may downregulate the expression of AR in LNCaP cells,inhibit their migration and proliferation abilities,and its mechanism of action may be related to the decreased expression of Bcl-xL and PCNA.