IGF-1R基因通过调控BMP2表达影响SMMC7721细胞的增殖和凋亡
发布时间：2019-03-16 11:52
【摘要】：背景与目的:胰岛素生长因子-1(insulin-like growth factor-1,IGF-1)是一种重要的肽类激素,通过与其受体(IGF-1 receptor,IGF-1R)和胰岛素受体(insulin receptor,IR)结合并激活下游信号通路发挥生物学作用,骨形态发生蛋白(bone morphogenetic proteins,BMPs)因可促进多种肿瘤细胞的增殖和侵袭而日益成为肿瘤分子研究领域的热点。该研究通过RNA干扰(RNAi)技术沉默IGF-1R基因,探讨其对肝癌SMMC7721细胞中BMP2表达的影响以及对细胞增殖和凋亡的影响。方法:构建靶向IGF-1R基因的RNAi真核表达质粒,转染至肝癌SMMC7721细胞,用反转录聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)和蛋白[质]印迹法(Western blot)检测IGF-1R和BMP2基因表达抑制效应,MTT实验检测IGF-1R基因沉默后细胞生长曲线,流式细胞术检测IGF-1R基因沉默后细胞凋亡情况。结果:成功构建靶向IGF-1R基因的真核表达质粒,IGF-1R-si RNA-1和IGF-1R-si RNA-2转染至肝癌SMMC7721细胞后,对IGF-1R基因的m RNA抑制效率分别达到68.9%和80.7%(P0.05),对BMP2基因的m RNA抑制效率分别达到79.5%和83.3%(P0.05),对IGF-1R蛋白表达抑制率分别为46.1%和62.1%,对BMP2蛋白表达抑制率分别为42.5%和60.9%(P0.05)。根据MTT实验结果,绘制的生长曲线显示,IGF-1R基因沉默后SMMC7721细胞增殖速率明显低于对照组(P0.05),细胞凋亡比例高于对照组(P0.05)。结论:IGF-1R基因沉默表达能介导BMP2基因不同水平下调表达,抑制SMMC7721细胞增殖,并促进细胞发生凋亡。
[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.
【作者单位】： 江西省九江市第三人民医院肿瘤科;江西省修水县第一人民医院肿瘤科;
【分类号】：R73-3
本文编号：2441296
[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.
【作者单位】： 江西省九江市第三人民医院肿瘤科;江西省修水县第一人民医院肿瘤科;
【分类号】：R73-3
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