發(fā)布時(shí)間：2019-03-16 11:52

【摘要】：背景與目的:胰島素生長(zhǎng)因子-1(insulin-like growth factor-1,IGF-1)是一種重要的肽類激素,通過(guò)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(hào)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(guò)RNA干擾(RNAi)技術(shù)沉默IGF-1R基因,探討其對(duì)肝癌SMMC7721細(xì)胞中BMP2表達(dá)的影響以及對(duì)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(cè)IGF-1R和BMP2基因表達(dá)抑制效應(yīng),MTT實(shí)驗(yàn)檢測(cè)IGF-1R基因沉默后細(xì)胞生長(zhǎng)曲線,流式細(xì)胞術(shù)檢測(cè)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(duì)IGF-1R基因的m RNA抑制效率分別達(dá)到68.9%和80.7%(P0.05),對(duì)BMP2基因的m RNA抑制效率分別達(dá)到79.5%和83.3%(P0.05),對(duì)IGF-1R蛋白表達(dá)抑制率分別為46.1%和62.1%,對(duì)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(zhǎng)曲線顯示,IGF-1R基因沉默后SMMC7721細(xì)胞增殖速率明顯低于對(duì)照組(P0.05),細(xì)胞凋亡比例高于對(duì)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(hào)】：R73-3

【相似文獻(xiàn)】

相關(guān)期刊論文 前3條

1 尉輝杰;劉麗;何堅(jiān);鄧月娥;孔凡靜;田玉慧;;蜂膠黃酮對(duì)大鼠顱骨成骨細(xì)胞增殖分化及BMP2表達(dá)的影響[J];中國(guó)現(xiàn)代醫(yī)學(xué)雜志;2009年19期

2 張喜善;蔡國(guó)棟;李建民;楊明峰;;頸椎不穩(wěn)黃韌帶TGFβ1、BMP2表達(dá)的實(shí)驗(yàn)研究[J];山東大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2007年12期

3 ;[J];;年期

相關(guān)會(huì)議論文 前2條

1 孔祥英;林娜;陳衛(wèi)衡;;不同治則方藥對(duì)激素性股骨頭壞死雞股骨頭內(nèi)TGF－β1和BMP2表達(dá)的影響[A];中華中醫(yī)藥學(xué)會(huì)骨傷分會(huì)第四屆第三次學(xué)術(shù)年會(huì)暨國(guó)家中醫(yī)藥管理局“十一五”重點(diǎn)�？疲▽２。┙ㄔO(shè)骨傷協(xié)作組經(jīng)驗(yàn)交流會(huì)論文匯編[C];2008年

2 孫立;田曉濱;張宇坤;郜勇;楊述華;;人BMP2、VEGF165雙基因真核表達(dá)質(zhì)粒的構(gòu)建及體外表達(dá)[A];2006年貴州省醫(yī)學(xué)會(huì)骨科學(xué)分會(huì)學(xué)術(shù)年會(huì)論文匯編[C];2006年

相關(guān)碩士學(xué)位論文 前4條

1 潘建俸;載BMP2質(zhì)粒DNA的殼聚糖溫敏水凝膠對(duì)牙周膜成纖維細(xì)胞相容性的評(píng)價(jià)[D];青島大學(xué);2015年

2 陳佳濱;BMP2和EGFP重組腺病毒體外轉(zhuǎn)染兔骨髓間充質(zhì)干細(xì)胞的研究[D];桂林醫(yī)學(xué)院;2014年

3 李勇;大鼠骨髓間充質(zhì)干細(xì)胞生物學(xué)特性及BMP2在誘導(dǎo)其向神經(jīng)元樣細(xì)胞分化中的作用[D];第三軍醫(yī)大學(xué);2004年

4 王卓;幼齡水貂毛囊發(fā)育及皮膚中β-catenin和BMP2表達(dá)規(guī)律的研究[D];中國(guó)農(nóng)業(yè)科學(xué)院;2014年

，

本文編號(hào)：2441296