摘要
目的探讨C-FLIP siRNA对大肠癌细胞株SW480凋亡的影响。方法常规方法培养大肠癌SW480细胞,待细胞处于对数生长期时,进行转染,分别转染C-FLIP siRNA重组体(观察组)和nonsilencing空载体(NC组),转染后24 h,将细胞传至96孔板中,两组转染细胞各传32孔,每8孔为一组平行孔,分别检测16 h、24 h、48 h、72 h细胞的增殖情况,并检测两种转染细胞中GHR受体mRNA和蛋白的表达情况。结果 C-FLIP siRNA重组体转染率达75%;大肠癌SW480培养后各时间点的吸光度值相比,差异有统计学意义(F=7.329,P=0.016),观察组的吸光度值均小于对照组,并且随着时间的延长,观察组吸光度值增加越来越慢,直至不增加;两组GHR受体蛋白水平相比,对照组高于观察组,差异有统计学意义(t=73.140,P=0.000);GHR mRNA表达观察组高于对照组,差异有统计学意义(t=12.685,P=0.000)。结论 C-FLIP siRNA重组体能够有效抑制大肠癌细胞株中SW480的增殖。
Objective To investigate the effect of C-FLIP siRNA on apoptosis of human coloreetal cancer cell line SW480. Methods Colorectal cancer SW480 cells were cultured. When cells were in the logarithmic growth phase, eolorectal cancer SW480 cells were transfected with C-FLIP siRNA recombinant and nonsilencing respectively （NC） vector. Proliferation of 16 h, 24 h, 48 h, 72 h cells were detected, respectively. The expression of GHR mRNA and protein in two kinds of transfected cells were detected. Results The construction of RNAi recombinant plasmid siRNA rate was 75% ; absorbance value at each time point after SW480 euhure in colorectal cancer was significantly different （F =7. 329, P =0. 016） , absorbance value in the observation group was smaller than that in the NC group. With the extension of time, the absorbance value in obinservation group was increased more slowly, finally had no increasing trend ; expression of GHR protein had significant difference between two groups （t = 73. 140, P = 0. 000） , the expres- sion of GHR mRNA had significant difference between two groups （t = 12. 685, P = 0. 000）. Conclusion The con- struction of C-FLIP siRNA recombinant can effectively inhibit the colorectal cancer cell lines SW480 in vitro.
出处
《胃肠病学和肝病学杂志》
CAS
2014年第8期967-969,共3页
Chinese Journal of Gastroenterology and Hepatology
