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目的:探讨PRL-3在人肝内胆管癌侵袭转移中的作用。方法:利用小RNA技术干扰肝内胆管癌细胞株PRL-3表达,并采用细胞划痕实验和Transwell体外侵袭实验评价PRL-3对肝内胆管癌细胞侵袭转移能力的影响。结果:RT-PCR和Western blot结果均显示转染PRL-3特异性siRNA-2组PRL-3表达明显降低(P<0.05)。PRL-3 siRNA-2组在划痕培养24h后划痕区域宽度占初始划痕区域宽度的百分比为(62.12±6.28)%,阴性对照组为(23.88±2.55)%,空白对照HCCC-9810组为(21.20±6.07)%。PRL-3siRNA-2组细胞的划痕两端距离相比明显较宽,分别与阴性对照组细胞和空白对照HCCC-9810组细胞相比均有统计学意义(P<0.05),后两者无显著性差异(P>0.05)。Transwell体外侵袭实验结果显示,PRL-3 SiRNA-2组细胞侵袭能力明显减弱,穿膜细胞数为(19.40±2.30)个/HP,明显少于阴性对照组(64.00±2.73)个/HP和正常HCCC-9810组(67.20±3.11)个/HP,差异有显著性(P<0.05);正常HCCC-9810组和阴性对照组无明显差异(P>0.05)。结论:PRL-3特异性siRNA能够抑制肝内胆管癌细胞HC-CC-9810中内源性PRL-3的表达,并可以明显抑制肝内胆管癌细胞的迁移侵袭能力。
Objective: To investigate the role of PRL-3 in invasion and metastasis of human intrahepatic cholangiocarcinoma. Methods: Small interfering RNA (RNAi) was used to interfere with the expression of PRL-3 in intrahepatic cholangiocarcinoma cell line. The effects of PRL-3 on the invasion and metastasis of intrahepatic cholangiocarcinoma cells were evaluated by cell scratch assay and Transwell invasion assay. Results: The results of RT-PCR and Western blot showed that the expression of PRL-3 in PRL-3-specific siRNA-2 group was significantly decreased (P <0.05). The percentage of the width of the scratched area to the width of the initial scratched area was (62.12 ± 6.28)% in the PRL-3 siRNA-2 group and (23.88 ± 2.55)% in the negative control group, respectively, while that in the blank control group HCCC-9810 group (21.20 ± 6.07)%. Compared with the cells in the negative control group and the blank control group, the cells in the PRL-3siRNA-2 group were significantly wider than those in the HCCC-9810 group (P <0.05), and the latter two had no significant difference Significant difference (P> 0.05). Transwell in vitro invasion assay showed that the invasion ability of PRL-3 SiRNA-2 group was significantly weakened, the number of transmembrane cells was (19.40 ± 2.30) / HP, significantly less than that of the negative control group (64.00 ± 2.73) / HP and normal HCCC-9810 group (67.20 ± 3.11) / HP, the difference was significant (P <0.05); there was no significant difference between normal HCCC-9810 group and negative control group (P> 0.05). Conclusion: PRL-3 specific siRNA can inhibit the expression of endogenous PRL-3 in intrahepatic cholangiocarcinoma cell line HC-CC-9810 and significantly inhibit the migration and invasion of intrahepatic cholangiocarcinoma cells.