【摘要】 目的探讨前列腺癌DU145细胞中解聚素—金属蛋白酶17(ADAM17)与转化生长因子α(TGF-α)活化的关系。方法培养前列腺癌DU145细胞,将细胞分为对照组、NC组、ADAM17-siRNA组、联合组。ADAM17-siRNA组、联合组均转染ADAM17 siRNA,NC组转染非ADAM17相关的阴性序列siRNA,对照组不转染。联合组转染72 h后加入TGF-α20μg/L干预。待NC组、ADAM17-siRNA组转染72 h后,收集对照组、NC组、ADAM17-siRNA组细胞,采用Western blot法检测ADAM17、表皮生长因子受体(EGFR)、磷酸化EGFR(p EGFR)蛋白表达,采用ELISA法检测TGF-α蛋白水平。取对照组、ADAM17-siRNA组及联合组TGF-α干预15 min后的上清液,采用Western blot法检测EGFR、p EGFR蛋白表达。结果ADAM17-siRNA组ADAM17、p EGFR、TGF-α蛋白均低于对照组及NC组(P均<0.05)。对照组、NC组、ADAM17-siRNA组间EGFR表达差异无统计学意义(P均>0.05)。联合组p EGFR蛋白表达高于对照组及ADAM17-siRNA组(P均<0.05)。对照组、ADAM17-siRNA组、联合组间EGFR蛋白表达差异无统计学意义(P均>0.05)。结论前列腺癌DU145细胞中ADAM17的表达与TGF-α及EGFR有关,ADAM17可能通过活化TGF-α进而激活p EGFR表达。更多还原

【Abstract】 Objective To investigate the relationship between a disintegrin and metalloprotease 17( ADAM17) and transforming growth factor-α( TGF-α) activation in prostate cancer cells DU145. Methods We cultured the prostate cancer DU145 cells.Then,the cells were divided into the control group,NC group,ADAM17-siRNA1 group and the combination group. ADAM17 siRNA was transfected into both the ADAM17-siRNA1 group and combination group. The NC group was transfected with non-DAM17 related sequence siRNA. After the combination group was transfected for 72 h,20 g / L TGF-α was added. After 72-hour transfection,the cells of the control group,NC group and ADAM17-siRNA1 group were collected. Western blotting was used to detect the protein expression of ADAM17,epidermal growth factor receptor( EGFR) and p EGFR. ELISA was used to detect the level of TGF-α protein.We collected the supernatant intervened for15 mins of TGF-α from the control group,ADAM17-siRNA1 group and the combination group,and Western blotting was used to detect the protein expression of EGFR and p EGFR. Results The protein expression of ADAM17,p EGFR and TGF-α in the ADAM17-siRNA1 group was lower than that of the control group( all P <0. 05). There was no significant difference in EGFR expression between the control group,NC group and ADAM17-siRNA1 group( P >0. 05). The expression of p EGFR in the combination group was higher than that in the control group and the ADAM17-siRNA1 group( P <0. 05). There was no significant difference in the EGFR expression between the control group,ADAM17-siRNA1 group and the combination group( P >0. 05). Conclusion The expression of ADAM17 in prostate cancer DU145 cells is correlated with the activation of TGF-α and EGFR,and ADAM17 activates p EGFR expression by activating TGF-α.更多还原