过表达SOX7对胆囊癌GBC-SD细胞增殖生物学特性的影响及其机制研究

Effects of SOX7 overexpression on proliferation and other biological characteristics of gallbladder carcinoma GBC-SD cells and its mechanism

目的探讨过表达SOX7(Sex determining region Y-box 7)基因对胆囊癌GBC-SD细胞株增殖、凋亡的影响及其可能涉及的细胞信号通路。方法用特异性的重组质粒pc DNA3.1-SOX7作为实验组转染GBC-SD细胞建立稳定过表达SOX7基因的细胞,空载体pc-DNA3.1-mock作为对照组。采用Edu细胞荧光染色法检测过表达SOX7基因对GBCSD细胞增殖的影响,同时采用Hoechst 33342细胞染色法检测GBC-SD细胞的凋亡。Western blot检测相关信号通路蛋白的表达。结果实验组中SOX7 mRNA的相对表达量为(353.0±28.1)%,相对于对照组(100.0±2.3)%表达明显增加,差异具有统计学意义(P<0.05)。对照组细胞在450 nm吸光度值为(4.6±0.4),较实验组(2.4±0.2)明显升高,差异具有统计学意义(P<0.05)。Edu免疫荧光染色结果表明,对照组细胞增殖比例(33.3±3.4)%高于实验组(12.2±4.2)%,差异具有统计学意义(P<0.05)。Hoechst 33342细胞凋亡荧光染色结果表明,对照组细胞凋亡比例(5.2±1.3)%较实验组(10.2±2.4)%降低,差异具有统计学意义(P<0.05)。Western blot实验结果表明,对照组PTEN蛋白的相对表达量为(0.2±0.02),而实验组的相对表达量为(0.7±0.04),对照组磷酸化Akt的相对表达量为(0.8±0.03),而实验组为(0.4±0.02),差异具有统计学意义(P<0.05)。结论过表达SOX7基因能显著抑制胆囊癌细胞GBC-SD的增殖,导致其凋亡增加。PTEN/Akt信号通路的激活可能与之有关。

Objective The aims of the study were to explore effects of SOX7（ Sex determining region Y-box 7） overexpression on proliferation and apoptosis of gallbladder carcinoma of GBC-SD cells and the possible signal pathways involved. Methods The specific recombinant plasmid-pc DNA3. 1-SOX7 was used as the experimental group to transfect GBC-SD cells and then establish a stable over-expressed SOX7 cell line. The empty vector pc-DNA3. 1-mock was used as a control group. Effects of SOX7 overexpression on the proliferation of GBC-SD cells were detected by Edu cell immunofluorescent staining assay. The apoptosis of GBC-SD cells were detected by Hoechst 33342 staining assay. The phosphorylation of Akt and PTEN protein related to signaling pathways were examined by Western blot. Results The relative expression of SOX7 mRNA in the experimental group was（ 353 ±28. 1） %,which was significantly increased when compared to the control group（ 100 ± 2. 3） %（ P 0. 05）. The absorbance at 450 nm in the control group was 4. 6 ± 0. 4,which was significantly higher than that in the experimental group（ 2. 4 ± 0. 2）（ P 0. 05）. The proportion of cell proliferation in the control group（ 33. 3 ± 3. 4） % was higher than that in the experimental group（ 12. 2 ± 4. 2） % in GBC-SD cells. They showed a statistically significance（ P 0. 05）. The proportion of apoptosis in the experimental group（ 10. 2 ±2. 4） % was higher than that in the control group（ 5. 2 ± 1. 3） % in GBC-SD cells. They also showed a statistically significance（ P 0. 05）. The relative expression level of PTEN protein in the experimental group（ 0. 7 ± 0. 04） significantly increased in GBC-SD cells in comparison with the control group（ 0. 2 ± 0. 02）（ P 0. 05）. The relative expression of phosphorylated Akt protein in the experimental group（ 0. 4 ± 0. 02） significantly decreased in GBC-SD cells when compared to the control group（ 0. 8 ± 0. 03）（ P 0. 05）.Conclusion The overexpression of SOX7 significantly inhibits the proliferation and induces apoptosis of GBC-SD cells. Activation of the PTEN/Akt signaling pathway may involve in this process.