干扰素α-1b联合环氧化酶-2抑制剂对肾癌细胞及其裸鼠移植瘤的抑制作用

Inhibitory effect of interferon α-1b and cyclooxygenase-2 inhibitor on renal cancer cell and xenografts in nude mouse

目的探讨干扰素α-1b(interferonα-1b,IFNα-1b)联合环氧化酶-2(cyclooxygenase-2,COX-2)抑制剂对人肾癌ACHN细胞及其裸鼠肾癌移植瘤生长的抑制作用及其相关机制。方法 ACHN细胞及裸鼠根据用药不同分为IFNα-1b组、NS398组(COX-2抑制剂)、联合用药组及空白对照组。CCK8检测用药24 h和48 h后各组细胞增殖抑制率;Western blot检测用药前后各组细胞bcl-xl、COX-2蛋白表达变化。测量并记录各组裸鼠移植瘤的大小,免疫组化方法检测裸鼠移植瘤组织中VEGF的表达。结果 2种药物对ACHN细胞均有抑制作用,且呈剂量依赖性,联合用药优于单用(P<0.05);IFNα-1b和NS398均能促进肿瘤细胞凋亡,联合用药促凋亡效果优于单用(P<0.05);Western blot结果显示用药各组的bcl-xl、COX-2蛋白表达与对照组相比均下调,联合用药的抑制效果优于单用(P<0.05)。免疫组化显示用药各组的裸鼠移植瘤中VEGF均受抑制,联合组抑制作用最明显(P<0.05)。结论 IFNα-1b联合COX-2抑制剂对ACHN细胞增殖及其移植瘤生长具有一定的抑制作用。

Objective To investigate the inhibitory effect on human ACHN cell line and its mice xenograft by using interferon α-1b combined with cyclooxygenase-2 inhibitor and the relevant mechanism in vitro and vivo experiment. Methods ACHN cell and the xenograft mice were devided into 4 groups（ IFN-α1b,NS398,IFNα-1b ＋ NS398 and control group）. The inhibitory effects were tested by CCK8（ Cell Counting Kit 8） assay after AHCN were treated for 24 h and 48 h. The expression of bcl-xl and COX-2 were detected by Western blot. The volume of the xenografts of ACHN cell line and testing the expression of VEGF in xenografts were measured by immunohistochemistry assay. Results Both IFNα-1b and NS398 exerted inhibitory effects on ACHN and this effects showed a rising trend with a increasing concentration of drugs. The combined group was more significant than monotherapy group（ P〈0. 05）. Western blot assay showed that IFNα-1b and NS398 downregulated the expression of bcl-xl and COX-2 in ACHN. The combined group was more significant than monotherapy group（ P〈0. 05）.The combined group has the greatest inhibitory effects on the xenografts of ACHN cell line compared with monotherapy group and control group（ P〈0. 05）. The expression of VEGF in tumor was obiviously inhibited in combined group compared with monotherapy group and control group（ P〈0. 05）. Conclusion IFNα-1b combined with NS398 can inhibit the proliferation of ACHN and suppress the tumor growth.