摘要
目的分析Jak1-Tyk2活性缺陷与人肾细胞癌中α-干扰素(IFN-α)抵抗性的相关性。方法使用ACHN、Caki-1、Caki-2、NC65等肾细胞癌株,通过WST-1进行细胞增殖实验后提取细胞蛋白,构建质粒并转染肾癌细胞株,提取细胞蛋白后进行分析。观察肾癌细胞对IFN-α的敏感性;IFN-α对肾癌细胞株中Jak1-Tyk2的磷酸化影响;增强Jak1-Tyk2表达对肾癌细胞对IFN-α耐药性的影响。结果 IFN-α对ACHN、Caki-1、NC65肾癌细胞株生长抑制作用呈现剂量依赖性,但不能有效抑制Caki-2细胞的生长。IFN-α处理后,ACHN细胞中Jak1,Tyk2及Stat1的表达水平未发生变化,但三者磷酸化水平显著增高,而在Caki-2细胞中未检测到Jak1及Tyk2的表达。IFN-α处理Jak1或Tyk2载体转染后的Caki-2中,Jak1、Tyk2和Stat1磷酸化显著增强。在过表达Jak1或Tyk2载体的Caki-2细胞中,该细胞对IFN-α的敏感性显著增强,呈剂量依赖性。结论 Jak1-Tyk2缺陷激活与肾癌对IFN-α的耐药有关,恢复Jak1和Tyk2的表达水平有助于逆转肾癌对IFN-α的耐药性。
Objective To analyze the correlation between Jak1-Tyk2 activity defect and interferon alpha resistance in human renal cell carcinoma. Methods Renal cell carcinomas such as ACHN,Caki-1,Caki-2,and NC65 were used to perform cell proliferation assays using WST-1 to extract cellular proteins. Then construct plasmids and transfect renal cell carcinoma cells,followed by extraction of cellular proteins for analysis. Results The growth inhibitory effect of IFN-α on ACHN,Caki-1 and NC65 renal cancer cell lines was dose-dependent,but it could not effectively inhibit the growth of Caki-2 cells. After treated by IFN-α,the expression levels of Jak1,Tyk2,and Stat1 in ACHN cells did not change,but the phosphorylation level of the three cells was significantly increased. However,no expression of Jak1 and Tyk2 was detected in Caki-2 cells. Phosphorylation of Jak1,Tyk2,and Stat1 was significantly enhanced in Caki-2 after transfection with the Jak1 or Tyk2 vector.In Caki-2 cells over-expressing Jak1 or Tyk2 vectors,the sensitivity of the cells to IFN-α was significantly enhanced in a dose-dependent manner. Conclusion The activation of Jak1-Tyk2 is related to the resistance of renal cell carcinoma to IFN-α. Recovering the expression levels of Jak1 and Tyk2 can reverse the resistance of renal cell carcinoma to IFN-α.
作者
毕如玫
胡新一
尚东浩
BI Ru-mei;HU Xin-yi;SHANG Dong-hao(Department of Urology,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,Chna.)
出处
《临床和实验医学杂志》
2018年第15期1575-1578,共4页
Journal of Clinical and Experimental Medicine
基金
国家自然科学基金(编号:81572502)
北京市医院管理局临床医学发展专项经费资助(编号:ZYLX201604)
