腺病毒介导RA538及反义c-myc在不同细胞系中作用及其机制

Transduction efficiency, biologic effects and mechanism of recombinant RA538, antisense C-myc adenovirus on different cell lines

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。

AIM To compare the transduction efficiency, biological and molecular mechanism of recombinant RA538, antisense c-myc adenovirus on different cell lines. METHODS Human gastric, esophageal cancer and 2BS cell line were treated with AV-RA 538, AV-ASc-myc or AV-LacZ. In order to compare their transduction efficiency, biological effects and molecular mechanism, X-gal stain, cellular morphology, MTT and RT-PCR analysis were used.RESULTS Adenovirus transduction efficiency determined by Recombinant LacZ adenovirus was found higher in SGC7901, EC109 and 2BS cell lines, but was lower in EC109 cell line. AV-RA538 and AV-ASc-myc could strongly inhibit cell growth and induce apoptosis of SGC7901 cells. The growth rates of the AV-RA538 or AV-ASc-myc-infected SGC7901 cells were inhibited by 76.3% and 44.1% respectively. The overexpression of RA538 and ASc-myc could down-regulate expression of c-myc, bcl-2 gene. AV-RA538 or AV-ASc-myc could not inhibit cell growth and induce apoptosis of EC109 and 2BS cells, and could not down-regulate expression of c-myc and bcl-2. CONCLUSION Adenovirus vector has highly transfering efficiency, and can mediate a high level of expression of interest gene in transducted cells, but the transduction efficiency of adenovirus is different on each cell line. AV-RA538 or AV-ASc-myc may inhibit growth and induce apoptosis of gastric cancer cell through high transduction efficiency of AV and inhibition of c-myc and be/-2 expression. AV-RAS38 and AV-ASc-myc can not inhibit cell growth and induce apoptosis of esophageal cancer and 2BS cell line, which may be related to transduction efficiency and regulation of expression gene on them.