Xia Jin-tang, Xu Bo, Wang Shao-cheng, Lai Yue-yuan, Wu Zhao-feng, Li Shu-hua, Li Wen
Department of Hepatobiliary Surgery, Guangzhou First Municipal People' s Hospital, Guangzhou Medical College, Guangzhou 510180, China.
Zhonghua Yi Xue Za Zhi. 2008 Jul 8;88(26):1846-50.
To evaluate the impact of the recombined adeno-associated virus encoding soluble tumor necrosis factor related apoptosis inducing ligand gene (rAAV-sTRAIL) on proliferation and apoptosis of human hepatocellular carcinoma (HCC) cells, and to investigate the feasibility and efficiency of transfection of rAAV-sTRAIL into human HCC cells by ultrasound microbubble intensifier.
Human HCC cells of the line HepG2 were transfected with rAAV-sTRAIL or rAAV-sTRAIL combined with microbubble echocontrast agent and appropriate dose of ultrasound irradiation. RT-PCR and Western blotting were used to detect the mRNA and protein expression of sTRAIL gene. MTT method was used to detect the proliferation inhibition rate, and the apoptosis rate of the HepG2 cells was evaluated by flow cytometry.
The expression levels of sTRAIL mRNA and protein were higher in the rAAV-TRAIL combined with ultrasound microbubble group than in the rAAV-sTRAIL group (both P < 0.05). The proliferation inhibition rate of the rAAV-TRAIL combined with ultrasound microbubble group was significantly higher than that of the rAAV-sTRAIL group (P < 0.05). The apoptotic effect of the rAAV-TRAIL combined with ultrasound microbubble group was greater than that of the rAAV-sTRAIL group (P < 0.05).
TRAIL has a potential role to inhibit e proliferation and induce apoptosis of human hepatocellular carcinoma cells. Ultrasound and microbubble echocontrast agent increase the transfection rate of rAAV vector into HCC cells.
评估编码可溶性肿瘤坏死因子相关凋亡诱导配体基因的重组腺相关病毒(rAAV-sTRAIL)对人肝癌(HCC)细胞增殖和凋亡的影响,并探讨超声微泡增强剂介导rAAV-sTRAIL转染人HCC细胞的可行性和效率。
用rAAV-sTRAIL或rAAV-sTRAIL联合微泡超声造影剂及适当剂量的超声照射转染人肝癌HepG2细胞系。采用RT-PCR和蛋白质印迹法检测sTRAIL基因的mRNA和蛋白表达。采用MTT法检测增殖抑制率,通过流式细胞术评估HepG2细胞的凋亡率。
rAAV-TRAIL联合超声微泡组sTRAIL mRNA和蛋白表达水平高于rAAV-sTRAIL组(均P<0.05)。rAAV-TRAIL联合超声微泡组的增殖抑制率显著高于rAAV-sTRAIL组(P<0.05)。rAAV-TRAIL联合超声微泡组的凋亡作用大于rAAV-sTRAIL组(P<0.05)。
TRAIL对抑制人肝癌细胞增殖和诱导凋亡具有潜在作用。超声和微泡超声造影剂可提高rAAV载体对HCC细胞的转染率。