RNA 干扰 Cofilin-1基因沉默对人肝癌细胞株 Huh-7侵袭和转移能力的影响

RNA 干扰 Cofilin-1基因沉默对人肝癌细胞株 Huh-7侵袭
和转移能力的影响
曹建平;龙晓兰;龚勇;李晓杰;谢海龙
【期刊名称】《医学研究生学报》
【年(卷),期】2015(000)005
【摘要】目的： Cofilin-1参与多种肿瘤的发病过程，但该基因在肝细胞肝癌中表达和作用不明确。

文中拟观察Cofilin-1基因在人肝癌组织中的表达情况，探讨下调Cofilin-1表达对肝癌细胞株Huh-7侵袭和转移能力的影响。

方法采用实时荧
光定量PCR检测Cofilin-1基因在肝癌组织和癌旁组织中的表达情况；体外合成Cofilin-1序列特异性小干扰RNA，应用脂质体转染Huh-7肝癌细胞株，实验分Cofilin-1-siRNA组（ Cofilin-1-siRNA转染Huh-7细胞）、非特异性-RNA组（ Ctrl-siRNA组，非特异性siRNA转染Huh-7细胞）、未转染组（未转染
Huh-7细胞），每组设2个平行孔。

Western blot 鉴定Cofilin-1蛋白水平变化。

细胞迁移及体外侵袭实验观察转染后细胞的侵袭能力的影响。

结果与癌旁组织相比，肝癌组织中Cofilin-1基因表达明显增强[（0．698±0．156） vs
（3．523±0．412）， P＜0．05]；Cofilin-1-siRNA组Cofilin-1的蛋白量为0．558±0．033，明显低于Ctrl-siRNA组（0．933±0．015）和未转染组（0．961±0．020），差异有统计学意义（P＜0．05）；体外细胞迁移和侵袭实验结果均显示：与Ctrl-siRNA组迁移及侵袭细胞数量[（79．00±1．33）个]和
未转染组[（74．50±1．35）个]比较，Cofilin-1-siRNA组[（58．50±1．78）个]明显降低（P＜0．05）；Cofilin-1-siRNA组穿膜Huh-7细胞数[（36．50±0．83）个]明显低于Ctrl-siRNA组[（60．20±1．60）个]与未转染
组[（51．50±1．14）个]，差异有统计学意义（P＜0．05）。

结论 Cofilin-1在肝癌组织中高表达，下调Cofilin-1基因的表达能抑制肝癌细胞株HuH-7的侵袭和转移能力。

%[Abstract ] Objective Cofilin-1 is involved in the pathogenesis of various tumours .However, the expression and effect of Cofilin-1 in hepatocellular carcinoma is not clear .The aim of this study is to observe the Cofilin-1 gene expression in human hepatocel-lular carcinoma (HCC) tissues, and to explore the effect of Cofilin-1 gene expression on invasion and metastasis of HCC HuH-7 cells. Methods Real-time quantitative PCR was used to assess the Cofilin-1 gene expression in human HCC tissues and normal tumor-ad-jacent tissues.The specific small interfering RNA ( siRNA) of Cofilin-1 sequence was synthetized in vitro , and was transfected into HCC HuH-7 cells using liposome transfection.The experiment was divided into Cofilin-1-siRNA group, Ctrl-siRNA group and un-transfected group.Western blot assay was used to detect the protein expression of Cofilin-1.Migration and invasion experiments in vitro were used to investigate the invasive ability of transfected cells. Results Compared with the adjacent liver tissue , Cofilin-1 gene ex-pression in human liver cancer tissue was significantly increased (0.698 ±0.156 vs
3.523 ±0.412, P<0.05).The expression of Cofilin-1 protein in Cofilin-1-siRNA group was 0.558 ±0.033, which was lower than that in Ctrl-siRNA group ( 0.933 ±0.015 ) <br> and in untransfected group (0.961 ±0.020) (P<0.05).The results of migration and invasion experiments in vitro showed that the amount of migration and invasion cells in Cofilin-1-siRNA group were significantly lower than Ctrl-siRNA group or untransfected group
(58.50 ±1.78 vs 79.00 ±1.33, 74.50 ±1.35,P<0.05; 36.50 ±0.83 vs 60.20
±1.60, 51.50 ±1.14, P<0.05). Conclusion Cofilin-1 is highly expressed in HCC, and the invasion and metastasis of HCC HuH-7 cells is suppressed by inhibiting the Cofilin-1 gene expression.
【总页数】5页(P465-469)
【作者】曹建平;龙晓兰;龚勇;李晓杰;谢海龙
【作者单位】421005 衡阳，湖南环境生物技术学院护理学院;421001 衡阳，南华大学肿瘤研究所;421001 衡阳，南华大学肿瘤研究所;423000 郴州，郴州市第一人民医院病理科;421001 衡阳，南华大学肿瘤研究所
【正文语种】中文
【中图分类】R735.7
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