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目的探讨神经胶质瘤细胞株SHG-44高迁移率族蛋白A1(HMGA1)基因表达下调后对吉西他滨化疗敏感性的影响。方法用HMGA1 siRNA慢病毒载体及阴性siRNA慢病毒载体分别转染SHG-44细胞。稳定转染HMGA1 siRNA的细胞为阳性组、稳定转染不含HMGA1的细胞为阴性组,未转染的细胞为对照组。RT-PCR和Western blot检测HMGA1 mRNA和蛋白的表达,MTT法和克隆集落形成实验检测吉西他滨对细胞生长、增殖的影响,流式细胞术检测细胞凋亡率。结果阳性组HMGA1 mRNA和蛋白的表达水平明显低于阴性组和对照组(P<0.05)。经吉西他滨处理后,MTT表明阳性组细胞生长抑制率显著高于阴性组,阳性组IC50为(0.279±0.013)μg/ml,明显低于阴性组的(0.746±0.020)μg/ml(P<0.05)。克隆形成实验表明阳性组克隆形成率显著低于阴性组(P<0.05),而细胞凋亡率显著高于阴性组(P<0.05)。结论神经胶质瘤细胞SHG-44中HMGA1基因沉默后,显著增强SHG-44细胞对吉西他滨的化疗敏感性。
Objective To investigate the effect of gemcitabine chemosensitivity on the expression of high mobility group box A1 (HMGA1) gene in glioma cell line SHG-44. Methods SHG-44 cells were transfected with HMGA1 siRNA lentiviral vector and negative siRNA lentivirus vector respectively. The cells stably transfected with HMGA1 siRNA were positive and the cells stably transfected with HMGA1 siRNA were negative. The untransfected cells were control. The expression of HMGA1 mRNA and protein was detected by RT-PCR and Western blot. The effect of gemcitabine on cell growth and proliferation was detected by MTT assay and colony formation assay. The apoptosis rate was detected by flow cytometry. Results The expression of HMGA1 mRNA and protein in the positive group was significantly lower than that in the negative group and the control group (P <0.05). After gemcitabine treatment, MTT showed that the growth inhibition rate of positive cells was significantly higher than that of negative cells (IC50 = 0.279 ± 0.013μg / ml, significantly lower than that of negative cells (0.746 ± 0.020μg / ml, P <0.05 ). The colony formation assay showed that the positive rate of colony formation in the positive group was significantly lower than that in the negative group (P <0.05), while the apoptosis rate was significantly higher than that in the negative group (P <0.05). Conclusion The silencing of HMGA1 gene in SHG-44 glioma cells significantly enhances the chemosensitivity of SHG-44 cells to gemcitabine.