构建稳定干扰RN181的RKO细胞系并研究其生长现象

Study of RN181 regulating the growth of RKO cell using an RN181 RKOKD recombinant cell line

目的构建稳定干扰RN181的重组RKO细胞系,并研究下调RN181水平后,重组细胞系在体内外生长的规律。方法 RN181干扰慢病毒感染RKO细胞系,荧光与Western blotting鉴定该重组细胞系,CCK-8法、平板克隆实验研究该重组细胞系在体外增殖规律,裸鼠成瘤实验研究其在体内增殖情况。结果重组细胞系荧光良好,Western blotting证实重组细胞干扰组与对照组的RN181表达量差异有统计学意义(P<0.05)。体外细胞生长曲线实验与平板克隆结果显示,干扰组与对照组之间增殖能力差异无统计学意义(P>0.05)。干扰组的瘤体生长速度明显低于对照组,差异有统计学意义(P<0.05)。结论成功构建稳定干扰RN181的肠癌细胞系,并证实了下调RN181对RKO细胞在体外生长无影响但在体内能显著抑制细胞生长。

Objective To establish RKO cell line stably expressing interfered RN181 and study the rhythm of down-regulation RN181 in vivo and in vitro. Methods Using RN181 knocking-down lentivirus to infect the RKO ceils. After identification by Fluorescence and Western blotting, CCK-8, platting clone and xenograft tumor assays were used to evaluate the effect of RN181 on RKO proliferation in vitro and in vivo. Results The protein levels of RN181 among the recombinant cell lines confirmed by Fluorescence and Western Blotting have significant differences （P〈0.05）. CCK-8 and platting clone assay have no significant differences between interference cell lines in vitro （P〉0.05）. Xenograft tumor have significant difference in growth speed between the two recombinant cell lines in vivo （P〈0.05）. Conclusion We have successfully constructed recombinant RKO cell lines with interfered RN181 expression. Down-regulation of RN181 could not influence the proliferation of RKO cell in vitro and in vivo.