【摘要】：背景與目的:胰島素生長因子-1(insulin-like growth factor-1,IGF-1)是一種重要的肽類激素,通過與其受體(IGF-1 receptor,IGF-1R)和胰島素受體(insulin receptor,IR)結(jié)合并激活下游信號通路發(fā)揮生物學(xué)作用,骨形態(tài)發(fā)生蛋白(bone morphogenetic proteins,BMPs)因可促進(jìn)多種腫瘤細(xì)胞的增殖和侵襲而日益成為腫瘤分子研究領(lǐng)域的熱點(diǎn)。該研究通過RNA干擾(RNAi)技術(shù)沉默IGF-1R基因,探討其對肝癌SMMC7721細(xì)胞中BMP2表達(dá)的影響以及對細(xì)胞增殖和凋亡的影響。方法:構(gòu)建靶向IGF-1R基因的RNAi真核表達(dá)質(zhì)粒,轉(zhuǎn)染至肝癌SMMC7721細(xì)胞,用反轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)(reverse transcription-polymerase chain reaction,RT-PCR)和蛋白[質(zhì)]印跡法(Western blot)檢測IGF-1R和BMP2基因表達(dá)抑制效應(yīng),MTT實(shí)驗(yàn)檢測IGF-1R基因沉默后細(xì)胞生長曲線,流式細(xì)胞術(shù)檢測IGF-1R基因沉默后細(xì)胞凋亡情況。結(jié)果:成功構(gòu)建靶向IGF-1R基因的真核表達(dá)質(zhì)粒,IGF-1R-si RNA-1和IGF-1R-si RNA-2轉(zhuǎn)染至肝癌SMMC7721細(xì)胞后,對IGF-1R基因的m RNA抑制效率分別達(dá)到68.9%和80.7%(P0.05),對BMP2基因的m RNA抑制效率分別達(dá)到79.5%和83.3%(P0.05),對IGF-1R蛋白表達(dá)抑制率分別為46.1%和62.1%,對BMP2蛋白表達(dá)抑制率分別為42.5%和60.9%(P0.05)。根據(jù)MTT實(shí)驗(yàn)結(jié)果,繪制的生長曲線顯示,IGF-1R基因沉默后SMMC7721細(xì)胞增殖速率明顯低于對照組(P0.05),細(xì)胞凋亡比例高于對照組(P0.05)。結(jié)論:IGF-1R基因沉默表達(dá)能介導(dǎo)BMP2基因不同水平下調(diào)表達(dá),抑制SMMC7721細(xì)胞增殖,并促進(jìn)細(xì)胞發(fā)生凋亡。
[Abstract]:Background & objective: insulin growth factor-1 (insulin-like growth factor-1,IGF-1) is an important peptide hormone through its receptor (IGF-1 receptor,IGF-1R) and insulin receptor (insulin receptor,. IR) binds and activates downstream signaling pathways to play a biological role. Bone morphogenetic protein (bone morphogenetic proteins,BMPs) has become a hot topic in the field of tumor molecular research because it can promote the proliferation and invasion of a variety of tumor cells. In this study, IGF-1R gene was silenced by RNA interference (RNAi) technique, and its effect on BMP2 expression, cell proliferation and apoptosis in hepatocellular carcinoma (SMMC7721) cells was investigated. Methods: Eukaryotic expression plasmid of RNAi targeting IGF-1R gene was constructed and transfected into SMMC7721 cells. Reverse transcription polymerase chain reaction (reverse transcription-polymerase chain reaction,) was used. The inhibitory effects of IGF-1R and BMP2 gene expression were detected by RT-PCR) and protein blotting (Western blot). The cell growth curve after IGF-1R gene silencing was detected by MTT assay, and the apoptosis after IGF-1R gene silencing was detected by flow cytometry. Results: the eukaryotic expression plasmid targeting IGF-1R gene was constructed successfully. IGF-1R-si RNA-1 and IGF-1R-si RNA-2 were transfected into SMMC7721 cells. The inhibition efficiency of m-RNA to IGF-1R gene was 68.9% and 80.7% respectively (P0.05), and the inhibition efficiency of m-RNA to BMP2 gene was 79.5% and 83.3% (P0.05), respectively. The inhibition rate of IGF-1R protein expression was 46.1% and 62.1%, and that of BMP2 protein expression was 42.5% and 60.9% respectively (P0.05). According to the results of MTT experiment, the growth curve showed that the proliferation rate of SMMC7721 cells after IGF-1R gene silencing was significantly lower than that of the control group (P0.05), and the percentage of apoptosis was higher than that of the control group (P0.05). Conclusion: the silencing expression of IGF-1R gene can mediate the down-regulation of BMP2 gene expression at different levels, inhibit the proliferation of SMMC7721 cells and promote the apoptosis of SMMC7721 cells.
【作者單位】： 江西省九江市第三人民醫(yī)院腫瘤科;江西省修水縣第一人民醫(yī)院腫瘤科;
【分類號】：R73-3

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