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TWEAK反义寡核苷酸对人结肠癌细胞系SW480 增殖及侵袭能力的影响
张延新* 赵红军 宋文刚 高凤兰 吕慧芳
解剖学报 ›› 2014, Vol. 45 ›› Issue (1) : 74-79.
TWEAK反义寡核苷酸对人结肠癌细胞系SW480 增殖及侵袭能力的影响
Effects of TWEAK antisense oligonucleotide on the proliferation and invasive capability of the human colon cancer cell line SW480
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 explore the effects of TNF-like weak inducer of apoptosis(TWEAK) antisense oligonucleotide on the proliferation and invasive capability of the human colon cancer cell line SW480. Methods Colon cancer SW480 cells were transfected with synthesized TWEAK antisense oligodeoxynucleotide (ASODN) and scrambled oligodeoxynucleotide (SCODN). The cells were divided into control group, LF control group, ASODN group and SCODN group; MTT was applied to detect the inhibition of tumor cell proliferation, flow cytometry to examine the cell cycle distribution, ELISA to investigate the expression of soluble TWEAK and its receptor Fn14 in supernatant of cell culture fluid, Western blotting and immunofluorescence cytochemistry(IF) to detect the expression of soluble TWEAK and its receptor Fn14 and Transwell invasion assay to observe the changes on invasive capability of colon cancer cells. Results There was no significant difference on proliferation between the control group, LF control group and SCODN group (P>0.05). Compared with the control group, the proliferation rate of ASODN group tumor cells was inhibited obviously (P<0.05) in a dosage-time dependent manner. The ratio of G2+M phase cells was remarkably higher than that of control group after transfection with TWEAK ASODN (P<0.05). The expression rates of TWEAK and its receptor Fn14 in both cell culture fluid and cells were lower than those of control group, and the difference was of statistical significance (all P<0.05). The inhibition rate of infiltration was 31.39% after transfection with TWEAK ASODN, higher than those of control groups (P<0.05). The expression of TWEAK and its receptor Fn14 in cell culture fluid and cells was closely correlated (all P<0.05). Conclusion TWEAK ASODN may affect the occurrence and development of tumor probably by inhibition of binding between TWEAK and its receptor Fn14, and inhibition of TWEAK protein expression may restrain the proliferation and infiltration of the human colon cancer cell line SW480.
肿瘤坏死因子样凋亡微弱诱导剂 / 结肠癌 / 增殖 / 浸润 / 四甲基偶氮唑盐法 / 流式细胞术 / 人
Tumor necrosis factor-like weak inducer of apoptosis / Colon cancer / Proliferation / Infiltration / MTT assay / Flow cytometry / Human
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2012年度河南省教育厅科学技术研究重点项目
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