Gelsolin表达下调降低鼠源肝癌细胞的侵袭能力  

Down-regulation of Geisolin Decreases the Invasive Ability of Mouse Heptocarcinoma Cell lines

作　　者：王绍清[1] 孙明忠[2] 刘淑清[3] 王波[1] 唐建武[1] 

机构地区：[1]大连医科大学病理学教研室,辽宁省大连市116044 [2]大连医科大学生物技术专业 [3]大连医科大学分子生物学教研室

出　　处：《中国肿瘤临床》2009年第16期948-952,共5页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金(编号:30572098);高等学校博士学科点专项科研基金资助(编号:0040161005)~~

摘　　要：目的:研究高低淋巴道转移力细胞株Hca-F(淋巴道转移>80%)和Hca-P(淋巴道转移<30%)细胞中gelsolin的表达及其对Hca-F细胞侵袭能力的影响,阐明gelsolin在肝癌淋巴道转移过程中发挥的作用方法:应用荧光差异双向凝胶电泳技术及质谱定量和鉴定gelsolin在Hca-F细胞和Hca-P细胞中的表达;构建pSileneer-shR-NA-gelsolin表达载体,稳定转染Hca-F细胞并获得gelsolin的表达稳定下调的Hca-F细胞株,同时建立稳定转染pSilencer-无关序列Hca-F细胞株作为对照,在mRN水平和蛋白水平检测干扰后gelsolin的表达;Transwell 小室体外侵袭试验观察下调gelsolin表达后对Hca-F 细胞侵袭能力的影响结果:Gelsolin基因在Hca-F细胞中表达高于是Hca-P细胞( F/P= 1.7 );获得gelsolin稳定下调的Hca-F细胞株,RT-PCR和Wesiern Blot分析结果表明在稳定转染Hca-F细胞中,gelsolin在基因水平和蛋白水平的表达明显下调;在体外侵袭实验中,Hca-F细胞、对照组细胞、干扰组细胞穿过人工基底膜的细胞在570nm处的吸光度分别为0.2007±0.0129、0.2236±0.0446和0.1557±0.0051;表明下调gelsolin蛋白表达后,干扰组细胞穿过人工基底膜的细胞吸光度显著降低(P<0.05)。下调Hca-F细胞中gelsolin的表达后,转移的肿瘤细胞数量减少,癌细胞的侵袭能力下降结论:通过抑制gelsoin基因和蛋白的表达,可以降低鼠源肝癌细胞Hca-F侵袭能力的作用。Objective： To observe the expression levels of gelsolin in mouse hepatocarcinoma cell lines [Hca-F （lymphatic metastasis〉80%） and Hca-P （lymphatic metastasis〈30%）], to investigate the influence of gelsolin expression on the invasion of Hca-F cells, and to explore the role of gelsolin in the lymphatic metasta- sis of mouse hepatocarcinoma. Methods： Fluorescent two-dimensional difference in-gel electrophoresis （2D -DIGE） and mass spectrometry were used to detect the expression of gelsolin in Hca-F and Hca-P cell lines. The gelsolin shRNAs and scrambled shRNAs sequences were constructed into the pSilencer-3.1 vector to get the pSilencer-shRNA-gelsolin vectors which would be transfected into Hca-F cells. The expressions of gelsolin at mRNA level and protein level in the stably-transfected cell lines were measured by RT-PCR and Western blot. The inhibitive effect of RNAi on the migration abilities of Hca-F cells were analyzed by Transwell units. Results： The expression level of gelsolin in Hca-F cell line was 1.7 times of that in Hca-P cell fine. The expression vector of pSilencer-shRNA-gelsolin was established and transfected into Hca-F cells successfully. In vitro invasion assay showed that the absorbences of the elution solutions for the cells passing the matrigel for the Hca-F group, the control group and the transfected group were 0.2007±0.0129, 0.2236±0.0446, and 0.1557 ± 0.0051 （P〈0.05）, respectively. The invasive ability of Hca-F cell line decreases in response to the down-regulation of gelsolin. Conclusion： The down-regulation of gelsolin at both the gene and protein levels can decrease the invasive abilities of mouse hepatocarcinoma cell line Hca-F, indicating that gelsolin might play an important role in the lymphatic metastasis of hepatocarcinoma.

关 键 词：GELSOLIN 荧光差异双向凝胶电泳 肝肿瘤 侵袭 淋巴道转移 

分 类 号：R735.7[医药卫生—肿瘤]