摘要
目的探讨选择性环氧化酶2(COX2)抑制剂抗结肠癌细胞株SW480与细胞周期蛋白D1(CyclinD1)、凋亡蛋白Bcl2的关系,明确COX2抑制剂抗结肠癌非COX2依赖性途径的细胞内分子机制。方法用逆转录聚合酶链反应(RTPCR)检测结肠癌细胞系SW480中COX2的mRNA表达水平;将选择性COX2抑制剂NS398,作用于结肠癌细胞系SW480,运用MTT法分别于0、24、48、72h检测细胞增殖状态;流式细胞仪观察NS398对细胞凋亡的影响,进一步采用Westernblot检测药物作用前后CyclinD1、Bcl2的表达。结果结肠癌细胞系SW480中未检测到COX2mRNA的表达;空白组S及G2/M期DNA百分含量与处理组比值分别为1.28、1.55(P<0.01),故NS398呈时间、剂量依赖性方式抑制SW480细胞增殖,促进其凋亡。同时,72h时空白组与NS398(75μmol/L)处理组CyclinD1、Bcl2表达水平比值分别为6.41和2.74(P<0.01),故两者表达水平随作用时间延长而下降。结论选择性COX2抑制剂NS398抗结肠癌存在着COX2非依赖性途径,可能通过CyclinD1,Bcl2影响结肠癌细胞SW480的增殖与凋亡,提示COX2抑制剂抗结直肠癌存在着新的靶点。
Objective To investigate the influence of selective COX 2 inhibitor NS 398 on proliferation and apoptosis of colorectal cancer and reveal the potential COX2 independent mechanism of NS 398 effect on colorectal cancer cell.Methods By using RT PCR assay,thye expression level of COX 2 mRNA in human colorectal cancer cell line SW480 was detected.After SW480 cells were treated with NS 398,MTT assay and flow cytometry were used to measure the proliferation and apoptosis in 0,24,48,72 h respectively.The expression of Cyclin D1 and Bcl 2 was detected by Western blot before and after treatment with NS 398.Results COX 2 mRNA expression was not detected in colorectal cancer cell line SW480.The ratios of S and G 2/M percentage,and Cyclin D1 and Bcl 2 expression in blank group and NS 398 treated group were 1.28 and 1.55 ( P <0.01),and 6.41 and 2.74 ( P <0.01),respectively.NS 398 inhibited the cells proliferation and induced apoptosis in a dose and time dependent manner and resulted in significant downregulation of CyclinD1 and Bcl 2.Conclusion NS 398 may inhibit the proliferation and induce apoptosis of colon cancer cell lines SW480 through down regulating the expression of Cyclin D1 and Bcl 2.The COX 2 independent effect did exist.It may be a new target of selective COX 2 inhibitor effect on colon cancer.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2004年第7期798-800,共3页
Chinese Journal of Experimental Surgery
基金
甘肃省教育厅科研项目(02309)
