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目的 比较重组RA538,反义c-myc及LacZ腺病毒(adenovirus,AV)对不同靶细胞的转染效率、生物学特性并探讨其作用的分子机制。 方法 以人胃癌细胞(SGC7901)、食管癌细胞(EC109)及人胚肺二倍体细胞(2BS)系为靶细胞,采用LacZ基因转染X-gal染色、形态学观察、MTT,RT-PCR等方法,研究重组RA538,反义c-myc及LacZ AV对上述细胞的转染效率,生物学作用及其分子机制。 结果 AV-LacZ进行重组腺病毒转导效率检测显示其对SGC7901,2BS细胞具有很高的转导效率,对EC109细胞转导效率较低。AV-RA538及AV-ASc-myc对SGC7901细胞能产生明显的生长抑制效应并诱导凋亡,其生长抑制率分别为76.3％和44.1％。AV-RA538及AV-ASc-myc对SGC7901细胞内源性c-myc,bcl-2基因的表达具有抑制作用。AV-RA538及AV-ASc-myc对EC109细胞及2BS细胞无明显的生长抑制及凋亡诱导作用,AV-RA538对EC109及2BS细胞中内源性c-myc,bcl-2基因的表达无调节作用。 结论 AV载体转导效率很高,能实现目的基因在转导细胞中的高水平表达,但对不同靶细胞的转染效率存在差别。AV-RA538,AV-ASc-myc对SGC7901的生长抑制及凋亡诱导作用可能是通过AV的高效转导及抑制c-myc,bcl-2的表达而实现的。AV-RA538,AV-ASc-myc对食管癌、2BS细胞系无类似作用可能与其对上述细胞的转导的
OBJECTIVE: To compare the transfection efficiency and biological characteristics of recombinant RA538, antisense c-myc and adenovirus (AV) to different target cells and to explore the molecular mechanism. Methods Human gastric cancer cells (SGC7901), esophageal cancer cells (EC109) and human embryonic lung diploid cells (2BS) lines were used as target cells. LacZ gene transfection was performed by X-gal staining. Morphological observation, MTT and RT- And other methods to study the transfection efficiency, biological effects and molecular mechanisms of recombinant RA538, antisense c-myc and LacZ AV on the above cells. Results AV-LacZ recombinant adenovirus transduction efficiency test showed that SGC7901,2BS cells with high transduction efficiency, EC109 cell transduction efficiency is low. AV-RA538 and AV-ASc-myc could significantly inhibit the growth of SGC7901 cells and induce apoptosis. The growth inhibition rates were 76.3% and 44.1%, respectively. AV-RA538 and AV-ASc-myc could inhibit the expression of endogenous c-myc and bcl-2 genes in SGC7901 cells. AV-RA538 and AV-ASc-myc had no obvious growth-inhibitory and apoptosis-inducing effects on EC109 cells and 2BS cells. No expression of endogenous c-myc and bcl-2 genes was observed in AV109 and 2BS cells by AV-RA538 effect. Conclusion The transduction efficiency of AV vector is high, which can achieve high level expression of target gene in transduced cells, but there are differences in transfection efficiency of different target cells. AV-RA538, AV-ASc-myc on SGC7901 growth inhibition and apoptosis induction may be through the efficient transduction of AV and inhibit c-myc, bcl-2 expression and achieve. AV-RA538, AV-ASc-myc on esophageal cancer, 2BS cell line similar effect may be its transduction of these cells