徐亦君1 张振玉1 徐娴1 曹伟军1 竺明晨2.人肝内胆管癌细胞PRL-3 的表达与侵袭转移研究[J].现代生物医学进展英文版,2012,12(24):4642-4646. |
人肝内胆管癌细胞PRL-3 的表达与侵袭转移研究 |
Expression of Phosphatase of Regenerating Liver-3 in IntrahepaticCholangiocarcinoma and its Role of Invasiveness and Metastasis* |
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DOI: |
中文关键词: 肝再生磷酸酶-3 RNA 干扰 肝内胆管癌 转移 |
英文关键词: PRL-3 siRNA Intrahepatic cholangiocarcinoma Metastasis |
基金项目:江苏省肿瘤医院青年创新研究项目(ZQ200904) |
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中文摘要: |
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HCCC-
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。 |
英文摘要: |
Objective: To evaluate the role of phosphatase of regenerating liver-3 in intrahepatic cholangiocarcinoma invasiveness
and metastasis. Methods: Human intrahepatic cholangiocarcinoma cell line, HCCC-9810 was transfected with PRL-3 siRNA. Scratch
wound healing assay and transwell chambers were used to evaluate the motllity and migrating velocity of cells. Results: The results of
PCR and western blot showed that expression of PRL-3 significantly decreased (P<0.05) after PRL-3 siRNA-2 transfection. Scratch
wound healing results showed that the ratio of wound distance 24h after scratching and original wound distance in PRL-3 siRNA-2 cells,
negative siRNA cells and parental HCCC-9810 cells, were respectively(62.12±6.28)%,(23.88±2.55)% and (21.20±6.07)%. The migrating
distances of PRL-3 siRNA-2 transfecting cells was signi覱cantly shorter than those of negative siRNA cells and parental HCCC-
9810 cells (P<0. 05). In invasion assay, the number of cells that had migrated into the lower chamber were (19.40 ±2.30)/HP in
PRL-3 siRNA-2 group,(64.00 ±2.73)/HP in negative control group and(67.20 ± 3.11)/HP in parental HCCC-9810 group. The difference
was significant between the group transfected with PRL-3 siRNA-2 and the other two groups (P<0.05). Conclusions: PRL-3 specific
siRNA can inhibit the expression of PRL-3 in HCCC-9810 cells and weaken the invasion and migration potency of HCCC-9810 cells. |
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