CAJ | 학술논문

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-1αmRNA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72 h后HIF-1α小干扰RNA纳米微粒组细胞的HIF-1αmRNA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-1αmRNA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.
배경:납미미립작위일충비교이상적비병독기인체송재체,구유무면역원성화치류성등우월성.이용납미기술화기인간우기술조단결양유도인자1α(hypoxia-inducible factor 1,HIF-1α)재식관린암조직적표체,강저암세포대화료약물적내약성,이론상성위능구억제식관암세포생장적유효방법.목적:탐토HIF-1α소간우RNA납미미립대인식관린암TE-1세포생장적억제작용.설계、시간급지점:이체외배양적식관린암TE-1세포위대상,기인수평완전수궤대조실험,우2007-01/2008-12재중산대학부속제삼의원중심실험실완성.재료:유상해생물공정공사합성HIF-1α소간우RNA,채용초성유화법제비납미미립.인식관린암TE-1세포주구자중과원상해세포고.방법:체외배양적인식관린암TE-1세포분위4조:분별위생리염수조,불함기인적납미미립조,HIF-1α소간우RNA조,HIF-1α소간우RNA납미미립조.주요관찰지표:RT-PCR검측HIF-1αmRNA적표체,Western-blot법검측HIF-1α단백적표체;류식세포의검측세포조망정황;MTT법검측세포증식정황.결과:처리72 h후HIF-1α소간우RNA납미미립조세포적HIF-1αmRNA표체화HIF-1α단백표체저우기타3조(P＜0.01),세포조망솔고우기타3조(P＜0.01).HIF-1α소간우RNA납미미립조세포증식수명현억제,생장완만,여기타3조비교,차이유현저성의의(P＜0.01).결론:HIF-1α소간우RNA납미미립능구유효감소식관린암TE-1세포적HIF-1αmRNA화단백적표체,제고종류세포적조망,현저억제TE-1세포적생장.
BACKGROUND: Nanosphere, an ideal nonviral gene delivery vector, is not excellence of immunogenicity and oncogenicity. Nanotechnology and gene interference are used to block hypoxia-inducible factor 1 alpha (HIF-1α) expression in esophageal squamous carcinoma tissue and decrease tolerance of malignant cells to chemotherapeutics. Theoretically, they become effective methods to inhibit malignant cell growth of esophageal squamous carcinoma. OBJECTIVE: To study the inhibitory effect of small interference RNA targeting HIF-1α (siRNA-HIF-1α) nanospheres on human esophageal squamous cancer TE-1 cell growth. DESIGN, TIME AND SETTING: Based on in vitro cultured esophageal squamous cancer TE-1 cells, a completely randomized controlled study was performed at the Central Laboratory, the Third Hospital Affiliated to Sun Yat-sen University from January 2007 to December 2008. MATERIALS: siRNA-HIF-1α was synthesized by Shanghai Bioengineering Company; siRNA-HIF-1α nanospheres were prepared using solvent evaporation technique; human esophageal squamous cancer TE cell strain was provided by Shanghai Cell Bank of the Chinese Academy of Sciences. METHODS: TE-1 cells cultured in vitro were assigned into four groups: saline, gene-free nanospheres, siRNA-HIF-1α, and siRNA-HIF-1α nanospheres groups. MAIN OUTCOME MEASURES: HIF-1α mRNA expression was detected by RT-PCR; HIF-1α protein expression was detected by Western blot; apoptosis of TE-1 cells was determined by flow cytometry; TE-1 cell growth was examined by MTT. RESULTS: At 72 hours after treatment, both HIF-1α mRNA expression and HIF-1α protein expression in the siRNA-HIF-1α nanospheres group were significantly less than other three groups (P < 0.01), but apoptotic rate was significantly greater than other three groups (P < 0.01). TE-1 cell growth was remarkably inhibited in the siRNA-HIF-1α nanospheres group, which was significantly different compared with other three groups (P < 0.01).CONCLUSION: siRNA-HIF-1α nanospheres can specifically reduce both HIF-1α mRNA and HIF-1α protein expressions in esophageal squamous carcinoma TE-1 cells, significantly increase tumor cell apoptosis, and remarkably inhibit TE-1 cell growth.