Department of Obstetrics and Gynecology, The Third Affiliated Hospital, Sun Yat-sen University, 600 TianHe Road, Guangzhou 510630, People's Republic of China.
Mol Cell Biochem. 2011 Jul;353(1-2):225-33. doi: 10.1007/s11010-011-0790-9. Epub 2011 Mar 26.
The type-1 insulin-like growth factor receptor (IGF-1R) is over-expressed by endometrial carcinoma, level of IGF-1R has been correlated with tumor progression, and high IGF-1R expression has been found to be an important prognostic factor. In the study, we used lentivirus-mediated shRNA targeting IGF-1R to silence its expression, then assessed the effect of down-regulation of this receptor on cell growth and chemosensitivity to cisplatin. Lentivirus-mediate shRNA was designed and transfected to the endometrial carcinoma HEC-1B cell. The IGF-1R mRNA and related protein expression, cell proliferation ability, cell apoptosis, and cell cycle change were detected. Cell proliferation inhibition rates, cell apoptosis, and level of cleaved caspase-9 were measured in various concentrations of cisplatin. The mRNA and protein level of IGF-1R, and the phosphorylated protein p-Akt, p-Erk were all suppressed after transfection. Cell proliferation was inhibited in successive five days after transfection, the highest inhibition rate was 43.28 ± 3.55% on day 5. After transfection, 24.96 ± 1.05% cells were in G(2)/M phase, and cell apoptotic rate increased from 10.66 ± 0.08 to 19.92 ± 1.34%. In various concentrations of cisplatin, transfected cells proliferation was significantly inhibited which made the IC50 value drop from 21.85 uM to 10.58 uM. Incubation with different concentrations of cisplatin for 48 h, cells apoptotic rate increased to 41.92 ± 2.5, 31.13 ± 2.76, 22.21 ± 4.63%, respectively, which was accompanied with increased cleaved caspase-9 expression. Lentivirus-mediated shRNA targeting IGF-1R has the potential to develop as a clinical treatment method in advanced and chemoresistant endometrial carcinoma.
1 型胰岛素样生长因子受体(IGF-1R)在子宫内膜癌中过度表达,IGF-1R 水平与肿瘤进展相关,高 IGF-1R 表达被认为是一个重要的预后因素。在本研究中,我们使用慢病毒介导的 shRNA 靶向 IGF-1R 沉默其表达,然后评估下调该受体对细胞生长和对顺铂化疗敏感性的影响。设计并转染慢病毒介导的 shRNA 至子宫内膜癌 HEC-1B 细胞。检测 IGF-1R mRNA 及其相关蛋白表达、细胞增殖能力、细胞凋亡和细胞周期变化。检测不同浓度顺铂对细胞增殖抑制率、细胞凋亡和 cleaved caspase-9 水平的影响。转染后 IGF-1R 的 mRNA 和蛋白水平以及磷酸化蛋白 p-Akt、p-Erk 均受到抑制。转染后连续 5 天抑制细胞增殖,第 5 天抑制率最高,为 43.28±3.55%。转染后 24.96±1.05%细胞处于 G2/M 期,细胞凋亡率从 10.66±0.08%增加到 19.92±1.34%。在不同浓度顺铂作用下,转染细胞增殖明显受到抑制,使 IC50 值从 21.85 μM 下降到 10.58 μM。不同浓度顺铂孵育 48 h 后,细胞凋亡率分别增加到 41.92±2.5、31.13±2.76、22.21±4.63%,同时 cleaved caspase-9 表达增加。靶向 IGF-1R 的慢病毒介导 shRNA 有可能成为晚期和耐药性子宫内膜癌的临床治疗方法。