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背景:纳米微粒作为一种比较理想的非病毒基因递送载体,具有无免疫原性和致瘤性等优越性。利用纳米技术和基因干扰技术阻断缺氧诱导因子1α(hypoxia-inducible factor 1,HIF-1α)在食管鳞癌组织的表达,降低癌细胞对化疗药物的耐药性,理论上成为能够抑制食管癌细胞生长的有效方法。目的:探讨HIF-1α小干扰RNA纳米微粒对人食管鳞癌TE-1细胞生长的抑制作用。设计、时间及地点:以体外培养的食管鳞癌TE-1细胞为对象,基因水平完全随机对照实验,于2007-01/2008-12在中山大学附属第三医院中心实验室完成。材料:由上海生物工程公司合成HIF-1α小干扰RNA,采用超声乳化法制备纳米微粒。人食管鳞癌TE-1细胞株购自中科院上海细胞库。方法:体外培养的人食管鳞癌TE-1细胞分为4组:分别为生理盐水组,不含基因的纳米微粒组,HIF-1α小干扰RNA组,HIF-1α小干扰RNA纳米微粒组。主要观察指标:RT-PCR检测HIF-1α mRNA的表达,Western-blot法检测HIF-1α蛋白的表达;流式细胞仪检测细胞凋亡情况;MTT法检测细胞增殖情况。结果:处理72h后HIF-1α小干扰RNA纳米微粒组细胞的HIF-1α mRNA表达和HIF-1α蛋白表达低于其他3组(P<0.01),细胞凋亡率高于其他3组(P<0.01)。HIF-1α小干扰RNA纳米微粒组细胞增殖受明显抑制,生长缓慢,与其他3组比较,差异有显著性意义(P<0.01)。结论:HIF-1α小干扰RNA纳米微粒能够有效减少食管鳞癌TE-1细胞的HIF-1α mRNA和蛋白的表达,提高肿瘤细胞的凋亡,显著抑制TE-1细胞的生长。
Background: Nanoparticles, as an ideal non-viral gene delivery vector, have the advantages of non-immunogenicity and tumorigenicity. Using nanotechnology and gene interference technology to block the expression of hypoxia-inducible factor 1α (HIF-1α) in esophageal squamous cell carcinoma and reduce the drug resistance of cancer cells to chemotherapeutic drugs, An effective method of cancer cell growth. Objective: To investigate the inhibitory effect of HIF-1α small interfering RNA on the growth of human esophageal squamous cell carcinoma TE-1 cells. DESIGN, TIME AND SETTING: The esophageal squamous cell carcinoma TE-1 cells cultured in vitro were completely randomized controlled and performed at the Central Laboratory of the Third Affiliated Hospital of Sun Yat-sen University from January 2007 to December 2008. MATERIALS: HIF-1α small interfering RNA was synthesized by Shanghai Bioengineering Co. and nanoparticles were prepared by phacoemulsification. Human esophageal squamous cell carcinoma TE-1 cell line purchased from the Shanghai Cell Bank of Chinese Academy of Sciences. Methods: Human esophageal squamous cell carcinoma TE-1 cells cultured in vitro were divided into 4 groups: saline group, gene-free nanoparticle group, HIF-1α small interfering RNA group and HIF-1α small interfering RNA nanoparticle group. MAIN OUTCOME MEASURES: The expression of HIF-1α mRNA was detected by RT-PCR, the expression of HIF-1α protein was detected by Western-blot, the apoptosis was detected by flow cytometry, and the proliferation was detected by MTT assay. Results: HIF-1α mRNA expression and HIF-1α protein expression in HIF-1α small interfering RNA nanoparticles group were lower than those in the other three groups (P <0.01) at 72 h after treatment, and apoptosis rate was higher than the other three groups (P < 0.01). HIF-1α small interfering RNA nanoparticles significantly inhibited cell proliferation, slow growth, compared with the other three groups, the difference was significant (P <0.01). Conclusion: HIF-1α small interfering RNA nanoparticles can effectively reduce the expression of HIF-1α mRNA and protein in esophageal squamous cell carcinoma TE-1 cells, increase the apoptosis of tumor cells and significantly inhibit the growth of TE-1 cells.