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    钙结合蛋白2基因介导胆管癌细胞增殖、转移和凋亡逃逸

    The CALB2 gene mediates the proliferation, metastasis and apoptosis escape of cholangiocarcinoma cells

      摘要
    • 摘要:
      目的： 探索钙结合蛋白2（calbindin 2，CALB2）在胆管癌中的表达和对胆管癌细胞的影响。
      方法： 免疫组织化学检测CALB2在胆管癌病人癌与癌旁样本中的表达。RT–PCR检测CALB2在胆管癌细胞系中表达和敲低CALB2载体的效率。细胞克隆、EDU和CCK–8实验检测各组增殖情况。伤口愈合实验和迁移实验检测敲低CALB2对胆管癌细胞转移能力的改变。APC单染法检测敲低CALB2对胆管癌细胞凋亡的影响。
      结果： 免疫组织化学结果显示，相较于癌旁组织，CALB2在胆管癌病人样本中高表达（P ＜ 0.05）。RT–PCR结果显示相较于HIBepic细胞，CALB2在RBE和QBC939细胞中高表达（P ＜ 0.05）。CCK–8、细胞克隆和EDU实验结果，相较于NC组，si–CALB2组抑制了RBE和QBC939细胞的增殖（P ＜ 0.05）。伤口愈合实验显示，相较于NC组，si–CALB2组的转移能力降低（P ＜ 0.05）。迁移实验结果显示，相较于NC组，si–CALB2组中迁移细胞数目减少（P ＜ 0.05）。随后细胞凋亡实验显示，相较于NC组，si–CALB2组细胞凋亡率升高（P ＜ 0.05）。
      结论： CALB2在胆管癌中高表达，且CALB2促进胆管癌细胞的增殖、转移和凋亡逃逸能力。

       

      Abstract:
      Objective To explore the expression of calbindin 2 (CALB2) in cholangiocarcinoma and its impact on cholangiocarcinoma cells.
      Methods Immunohistochemistry was used to detect the expression of CALB2 in cancerous and adjacent non-cancerous samples from patients with cholangiocarcinoma. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of CALB2 in cholangiocarcinoma cell lines and the efficiency of CALB2 knockdown vectors. Cell cloning, EdU, and CCK-8 assays were used to assess the proliferation of each group. Wound healing and migration assays were used to detect the changes in the metastatic ability of cholangiocarcinoma cells after CALB2 knockdown. Annexin V-FITC/PI staining was used to detect the effect of CALB2 knockdown on apoptosis of cholangiocarcinoma cells.
      Results Immunohistochemical results showed that CALB2 was highly expressed in cholangiocarcinoma patient samples compared to adjacent non-cancerous tissues (P ＜ 0.05). RT-PCR results indicated that CALB2 was overexpressed in RBE and QBC939 cells compared to HIBepic cells (P ＜ 0.05). CCK-8, cell cloning, and EdU assay results showed that the si-CALB2 group inhibited the proliferation of RBE and QBC939 cells compared to the NC group (P ＜ 0.05). Wound healing assays indicated that the metastatic ability of the si-CALB2 group was reduced compared to the NC group (P ＜ 0.05). Migration assay results showed that the number of migrated cells in the si-CALB2 group was decreased compared to the NC group (P ＜ 0.05). Subsequent apoptosis assays revealed that the apoptosis rate of cells in the si-CALB2 group was increased compared to the NC group (P ＜ 0.05).
      Conclusions CALB2 is highly expressed in cholangiocarcinoma and promotes the proliferation, metastasis, and apoptosis escape of cholangiocarcinoma cells.

       

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