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目的探讨原钙黏蛋白10(PCDH10)在胰腺癌细胞株中的表达情况以及生物学功能。方法利用反转录PCR检测PCDH10在CAPAN-1、PANC-1、ASPC-1、BXPC-3胰腺癌细胞以及HPDE6-C7人正常胰腺导管上皮细胞的表达情况。将质粒pc DNA3.1-PCDH10与质粒pc DNA3.1-vector通过脂质体转染至BXPC-3人胰腺癌细胞,转染后通过反转录PCR检测BXPC-3细胞中PCDH10 mRNA的水平、Western blot法检测其蛋白水平;CCK-8法和克隆形成实验检测细胞增殖情况,膜联蛋白Ⅴ-碘化丙啶双标记结合流式细胞术检测细胞凋亡,Transwell~(TM)侵袭和迁移实验、划痕实验检测BXPC-3细胞的侵袭和迁移。结果与正常胰腺导管上皮细胞相比较,PCDH10在CAPAN-1、PANC-1、BXPC-3胰腺癌细胞中表达明显下调。反转录PCR及Western blot法检测结果表明,转染pc DNA3.1-PCDH10的BXPC-3实验组细胞PCDH10表达明显高于转染pc DNA3.1-vector的对照组细胞。与对照组相比,过表达PCDH10后,BXPC-3细胞的增殖速度明显降低、克隆形成数明显减少;细胞凋亡率明显增加,细胞的侵袭、迁移能力明显降低。结论 PCDH10在胰腺癌细胞中表达下调,过表达PCDH10能够明显抑制BXPC-3胰腺癌细胞的增殖、侵袭和迁移能力,并且诱导BXPC-3细胞凋亡。
Objective To investigate the expression and biological function of PCDH10 in pancreatic cancer cell lines. Methods The expression of PCDH10 in normal pancreatic ductal epithelial cells of CAPAN-1, PANC-1, ASPC-1 and BXPC-3 pancreatic cancer cells and HPDE6-C7 human pancreatic ductal epithelial cells were detected by reverse transcription PCR. The pcDNA3.1-PCDH10 plasmid and pcDNA3.1-vector plasmid were transfected into BXPC-3 human pancreatic cancer cells by lipofectamine. The expression of PCDH10 mRNA in BXPC-3 cells was detected by reverse transcription PCR. Western blot was used to detect the protein level. Cell proliferation was detected by CCK-8 assay and clonogenic assay. Apoptosis, Transwell TM invasion and migration were detected by annexin Ⅴ-propidium iodide double labeling combined with flow cytometry The invasion and migration of BXPC-3 cells were detected by experiments and scratches. Results Compared with normal pancreatic ductal epithelial cells, the expression of PCDH10 was significantly down-regulated in CAPAN-1, PANC-1 and BXPC-3 pancreatic cancer cells. The results of reverse transcription PCR and Western blot showed that the expression of PCDH10 in BXPC-3 cells transfected with pcDNA3.1-PCDH10 was significantly higher than that in pcDNA3.1-vector transfected cells. Compared with the control group, the proliferation rate of BXPC-3 cells was significantly decreased after PCDH10 was overexpressed, and the number of clone formation was significantly decreased; the apoptosis rate of BXPC-3 cells was significantly increased, and the invasion and migration ability of BXPC-3 cells was significantly decreased. Conclusion The expression of PCDH10 is down-regulated in pancreatic cancer cells. Overexpression of PCDH10 can significantly inhibit the proliferation, invasion and migration of BXPC-3 pancreatic cancer cells and induce the apoptosis of BXPC-3 cells.