siRNA阻断NF-κB信号通路对食管鳞癌细胞增殖、耐药的影响

Knockdown of the NF-κB signaling pathway by small interference RNA inhibits the proliferation of esophageal squamous cancer cells

目的:通过RNA干扰(RNAi)阻断食管鳞癌细胞中NF-κB信号通路,研究其与肿瘤细胞增殖、耐药的关系。方法:使用NF-κB p65 siRNA分别转染人食管鳞癌Eca109和EC9706细胞72h,以未转染的Eca109和EC9706细胞为对照,采用Western blot检测p65蛋白的表达;MTT法检测转染NF-κBp65siRNA24h、48h、72h后Eca109和EC9706细胞的增殖情况及转染同时联合应用不同质量浓度5-Fu(0mg/L,16.35mg/L,32.7mg/L,327mg/L,3270mg/L,6540mg/L)对Eca109和EC9706细胞增殖的影响。结果:①NF-κB亚单位p65在Eca109和EC9706细胞质中高表达,p65siRNA可有效阻断p65蛋白表达。②MTT实验表明,转染组细胞存活率较未转染组明显下降(P<0.05);与化疗药5-Fu联用,转染组和未转染组细胞的增殖活性随着5-Fu浓度的增加有下降趋势,但在同一浓度,转染p65siRNA的细胞与未转染组相比,细胞增殖活性明显下降(P<0.05)。结论:应用RNAi技术可有效干扰p65的表达,抑制食管鳞癌细胞的增殖,增强对化疗药5-Fu的敏感性。因此,可将阻断NF-κB信号通路作为基因治疗的靶点。

Aim ： To investigate cell proliferation and resistance to chemotherapy of esophageal squamous cell carcinoma （ESCC） cells after inhibition of the NF-κB signaling pathway. Methods： NF-κB p65 siRNA was used to transfect Eca109 and EC9706 cells for 72 h, respectively. The untransfeeted Eca109 and EC9706 cells were used as control. The protein level of p65 was detected by Western blot. Cell viability was detected by MTT after the ESCC cells were transfccted with or without p65 siRNA for 24 h, 48 h, 72 h. The sensitivity of Eea109 and EC9706 to 5-Fu transfected with or without p65 siRNA was evaluated by MTT. Results： NF-κB p65 was expressed in the cytoplasm of 2 ESCC cell lines, p65 siRNA decreased the expression of p65 and inhibited the cell proliferation. Compared with the untransfected cells, the viability of 2 transfected ESCC cells was significantly suppressed at the same concentration of 5-Fu （ P 〈 0.05）. Conclusion： RNAi can effectively inhibit the expression of p65. p65 siRNA has anti-proliferative effects and increases the sensitivity of ESCC cells to 5-Fu, and therefore NF-κB might be a good target for cancer treatment.