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目的观察肿瘤坏死因子样凋亡微弱诱导剂(TWEAK)对人结肠癌细胞系SW480增殖及侵袭能力的影响。方法合成TWEAK反义寡核苷酸(ASODN)、错义寡核苷酸(SCODN)后转染人结肠癌细胞系SW480,将细胞分为空白对照组、脂质体(LF)对照组、ASODN组、SCODN组;采用MTT法检测肿瘤细胞增殖抑制情况,流式细胞术(FCM)检测肿瘤细胞周期分布,ELISA法检测细胞培养液上清中可溶性TWEAK及其受体成纤维细胞生长因子诱导早期反应蛋白14(Fn14)的表达,Western blotting法、免疫荧光细胞化学法(IF)检测细胞中的TWEAK及Fn14蛋白的表达,运用Transwell侵袭实验观测结肠癌细胞侵袭能力的变化。结果空白对照组、LF对照组、SCODN组之间结肠癌细胞的增殖情况无明显差异(P>0.05),与对照组相比ASODN组肿瘤细胞增殖受到明显抑制(P<0.05),且呈剂量-时间依赖关系;转染TWEAK ASODN后,G2+M期细胞比例明显高于对照组(P<0.05),TWEAK及其受体Fn14在结肠癌细胞培养液及细胞内的表达均低于对照组,差异均有统计学意义(P<0.05);转染TWEAK ASODN后肿瘤细胞侵袭抑制率为31.39%,明显高于对照组(P<0.05);TWEAK及其受体Fn14在结肠癌细胞培养液及细胞内的表达均密切相关(P<0.05)。结论 TWEAK ASODN可能通过抑制TWEAK与其受体Fn14结合干扰肿瘤的发生发展,抑制TWEAK表达能抑制人结肠癌细胞系SW480的增殖与侵袭。
Objective To observe the effect of tumor necrosis factor-like weak inducer (TWEAK) on the proliferation and invasion of human colon carcinoma cell line SW480. Methods Human colorectal cancer cell line SW480 was transfected with TWEAK antisense oligonucleotide (ASODN) and missense oligonucleotide (SCODN). The cells were divided into blank control group, lipofectamine (LF) control group, ASODN Group and SCODN group. The inhibition of tumor cell proliferation was detected by MTT assay. The cell cycle distribution was detected by flow cytometry (FCM). The soluble TWEAK and its receptor fibroblast growth factor-induced early stage The expression of Fn14 and Fn14 were detected by Western blotting and immunofluorescence cytochemistry (IF). The invasion ability of colon cancer cells was detected by Transwell invasion assay. Results There was no significant difference in the proliferation of colon cancer cells between blank control group, LF control group and SCODN group (P> 0.05). Compared with the control group, the proliferation of tumor cells in ASODN group was significantly inhibited (P <0.05) (P <0.05). The expression of TWEAK and its receptor Fn14 in colon cancer cell culture medium and cells were lower than that of the control group (P <0.05). The inhibitory rate of tumor cell invasion after transfection with TWEAK ASODN was 31.39%, which was significantly higher than that of the control group (P <0.05). TWEAK and its receptor Fn14 in colon cancer cell culture medium And intracellular expression were closely related (P <0.05). Conclusion TWEAK ASODN may interfere with the tumorigenesis by inhibiting the binding of TWEAK to its receptor Fn14, and inhibiting the expression of TWEAK may inhibit the proliferation and invasion of human colon cancer cell line SW480.