PLCD1对胰腺癌CAPAN-1和BXPC-3细胞上皮间充质转化的影响
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Effect of phospholipase PLCD1 on epithelial-mesenchymal transition of pancreatic cancer cells CAPAN-1 and BXPC-3
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    摘要:

    目的:进一步探索肿瘤相关基因磷脂酶Cδ1(phospholipase C δ1,PLCD1)对胰腺癌细胞系增殖、侵袭及迁移的影响和潜在分子行为机制。方法:用脂质体法将pcDNA3.1-PLCD1质粒转入肿瘤,同时设立空载体组;综合利用CCK-8、克隆形成实验、划痕实验和Transwell验证PLCD1对胰腺癌细胞增殖、迁移及侵袭的影响,Western blot实验研究其潜在机制。结果:瞬时转染过表达质粒后的胰腺癌细胞CAPAN-1和BXPC-3中,PLCD1表达明显提高(P<0.05);CCK-8和克隆形成实验结果表明,高表达的PLCD1抑制胰腺癌细胞增殖速度(P<0.05);划痕实验和Transwell实验结果表明,细胞运动、侵袭及迁移能力下降(P<0.05)。Western blot实验发现,MAPK/ERK通路中磷酸化的ERK 1/2,间质细胞标志分子N-cadherin和Vimentin表达受抑制(P<0.05),同时上皮细胞标志分子E-cadherin表达上调(P<0.05)。结论:本研究发现PLCD1可能通过抑制MAPK/ERK通路来抑制上皮间充质转化,从而抑制胰腺癌细胞的增殖、侵袭及迁移。本研究为进一步探索PLCD1在胰腺癌中的作用机理奠定了基础。

    Abstract:

    Objective:To investigate the effects of the phospholipase Cδ1(PLCD1),a potential tumour gene,in pancreatic cancer cells,on the proliferation,migration and invasion of pancreatic cancer cells. Methods:Liposome method was used to transfer the pcDNA3.1-PLCD1 to pancreatic cancer cells and the Vector group with empty vector was set. The proliferation,migratory and invasive abilities of pancreatic cancer cells were detected with CCK-8 assay,clone formation assay,Transwell assay and wound healing assay. In additional,Western blot was used to demonstrate the potential mechanism. Results:PLCD1 was significant up-regulated in CAPAN-1 and BX-PC-3 cells after being transiently transfected with the PLCD1 over-expressing plasmid(P<0.05). The ability of proliferation showed significant inhibition with CCK-8 and clone formation assay(P<0.05). The motility,invasion and migration were remarkable restrained with Transwell assay and wound healing assay(P<0.05). Additionally,Western blot showed PLCD1 induced MAPK/ERK pathway molecule p-ERK1/2(P<0.05),as well as the mesenchymal markers N-cadherin and Vimentin were decreased(P<0.05). And the ex-pression of the epithelial marker E-cadherin was increased(P<0.05). Conclusion:PLCD1 may inhibit the proliferation,migration and invasion of pancreatic cancer cells through inhibiting the MAPK/EKR pathway and suppressing the EMT,which lay the foundation for the further study of PLCD1 in pancreatic cancer.

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张桐,姜政. PLCD1对胰腺癌CAPAN-1和BXPC-3细胞上皮间充质转化的影响[J].重庆医科大学学报,2020,45(2):245-

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  • 在线发布日期: 2020-04-21
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