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LncRNA GHET1靶向miR-27b对胆囊癌细胞增殖、凋亡及转移的影响

Effect of LncRNA GHET1 on Proliferation, Apoptosis, and Metastasis of Gallbladder Cancer Cells by Targeting miR-27b

  • 摘要:
    目的 探讨GHET1对胆囊癌细胞生物学行为的影响及其对miR-27b的调控机制。
    方法 采用实时荧光定量PCR检测50例胆囊癌样本中GHET1与miR-27b的表达情况。利用si-NC载体、si-GHET1载体、miR-27b抑制物以及si-GHET1载体+miR-27b抑制物转染至SGC-996细胞,并分别作为对照组、GHET1干扰组、miR-27b干扰组以及GHET1+miR-27b干扰组。采用MTT法、流式细胞术以及Transwell实验检测各组细胞增殖、凋亡以及转移情况,以荧光素酶报告基因实验验证GHET1对miR-27b的调控作用。
    结果 GHET1在癌组织中的表达较癌旁组织升高,而miR-27b在癌组织中的表达较癌旁组织降低,GHET1与miR-27b表达呈负相关(P<0.05)。GHET1表达与TNM分期和淋巴结转移相关(P<0.05)。高GHET1表达与胆囊癌患者预后较差相关(P<0.05)。与对照组比较,GHET1干扰组细胞增殖能力降低、凋亡率升高且细胞转移数量减少,而miR-27b干扰组细胞增殖能力升高、凋亡率降低且细胞转移数量增加(P<0.05)。与GHET1干扰组比较,GHET1+miR-27b干扰组细胞增殖能力升高、凋亡率降低且细胞转移数量增加(P<0.05)。GHET1可以海绵吸附miR-27b并抑制其表达。
    结论 GHET1靶向miR-27b促进胆囊癌细胞增殖和转移并抑制细胞凋亡,提示GHET1/miR-27b轴在胆囊癌进展中具有一定的作用。

     

    Abstract:
    Objective To investigate the effect of GHET1 on the biological behavior of gallbladder cancer cells and the regulatory mechanism of GHET1 on miR-27b.
    Methods The expression of GHET1 and miR-27b in 50 samples of gallbladder cancer was detected by real-time quantitative PCR. The si-NC vector, si-GHET1 vector, miR-27b inhibitor, and si-GHET1 vector+miR-27b inhibitor were transfected into SGC-996 cells and set as the control group, GHET1 interference group, miR-27b interference group, and GHET1+miR-27b interference group. Cell proliferation, apoptosis, and metastasis in each group were detected by MTT, flow cytometry, and Transwell assays. The regulatory effect of GHET1 on miR-27b was validated by luciferase reporter gene assay.
    Results GHET1 expression was higher in cancer tissues than that in paracancerous ones. miR-27b expression was lower in cancer tissues than that in paracancerous tissues. GHET1 was negatively correlated with miR-27b expression (P<0.05), and GHET1 expression was associated with TNM staging and lymph node metastasis (P<0.05). High GHET1 expression was associated with poor prognosis of patients with gallbladder cancer (P<0.05). Compared with the control group, the GHET1 interference group showed decreased cell-proliferation ability, increased apoptosis rate, and reduced number of cell metastasis. The miR-27b interference group showed increased cell-proliferation ability, decreased apoptosis rate, and increased number of cell metastasis (P<0.05). Compared with the GHET1 interference group, the GHET1+miR-27b interference group showed increased cell-proliferation ability, decreased apoptosis rate, and increased number of cell metastasis (P<0.05). GHET1 inhibited miR-27b expression by acting as a sponge of miR-27b.
    Conclusion GHET1 promotes the proliferation and metastasis and inhibits the apoptosis of gallbladder cancer cells by targeting miR-27b, suggesting that GHET1/miR-27b axis plays a role in gallbladder cancer progression.

     

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