中南大学学报（医学版）
中南大學學報（醫學版）
중남대학학보（의학판）
JOURNAL OF CENTRAL SOUTH UNIVERSITY (MEDICAL SCIENCES)
2014年
12期
1221-1227
,共7页
肝细胞癌%BTB/POZ结构域蛋白7%Rho关联卷曲螺旋蛋白激酶2%法舒地尔%侵袭转移
肝細胞癌%BTB/POZ結構域蛋白7%Rho關聯捲麯螺鏇蛋白激酶2%法舒地爾%侵襲轉移
간세포암%BTB/POZ결구역단백7%Rho관련권곡라선단백격매2%법서지이%침습전이
hepatocellular carcinoma%BTB/POZ domain containing 7%Rho-associated,coiled-coil containing protein kinase 2%Fasudil%invasion and metastasis
目的：观察Rho激酶抑制剂法舒地尔(Fasudil)对人高侵袭潜能HCC细胞株3(human high metastatic liver cancer cells 3，HCCLM3)侵袭转移的影响，并且探讨其作用的机制。方法：应用100μmol/L Fasudil作用于HCCLM3细胞，采用肌动蛋白微丝荧光染色和侵袭小室实验观察HCCLM3细胞的运动侵袭能力。HCCLM3细胞经过处理后分为阴性对照组、Fasudil作用组、BTBD7干扰组，通过Western印迹检测BTB/POZ结构域蛋白7(BR-C, ttk and bab/pox virus domain containing 7，BTBD7)、Ras同系物家族成员C(ras homolog family member C，RhoC)、Rho关联卷曲螺旋蛋白激酶2(Rho-associated, coiled-coil containing protein kinase 2，ROCK2)、MMP2和MMP9蛋白表达水平，酶谱分析法检测MMP2和MMP9活性水平。BTBD7干扰组作为阳性对照。结果：Fasudil处理后HCCLM3侵袭运动能力下降，BTBD7，RhoC， ROCK2蛋白表达下调，MMP2和MMP9活性降低，与阴性对照组比较差异有统计学意义(均P<0.01)。结论：Fasudil具有干预BTBD7-ROCK2信号通路、抑制HCC侵袭转移的重要作用。
目的：觀察Rho激酶抑製劑法舒地爾(Fasudil)對人高侵襲潛能HCC細胞株3(human high metastatic liver cancer cells 3，HCCLM3)侵襲轉移的影響，併且探討其作用的機製。方法：應用100μmol/L Fasudil作用于HCCLM3細胞，採用肌動蛋白微絲熒光染色和侵襲小室實驗觀察HCCLM3細胞的運動侵襲能力。HCCLM3細胞經過處理後分為陰性對照組、Fasudil作用組、BTBD7榦擾組，通過Western印跡檢測BTB/POZ結構域蛋白7(BR-C, ttk and bab/pox virus domain containing 7，BTBD7)、Ras同繫物傢族成員C(ras homolog family member C，RhoC)、Rho關聯捲麯螺鏇蛋白激酶2(Rho-associated, coiled-coil containing protein kinase 2，ROCK2)、MMP2和MMP9蛋白錶達水平，酶譜分析法檢測MMP2和MMP9活性水平。BTBD7榦擾組作為暘性對照。結果：Fasudil處理後HCCLM3侵襲運動能力下降，BTBD7，RhoC， ROCK2蛋白錶達下調，MMP2和MMP9活性降低，與陰性對照組比較差異有統計學意義(均P<0.01)。結論：Fasudil具有榦預BTBD7-ROCK2信號通路、抑製HCC侵襲轉移的重要作用。
목적：관찰Rho격매억제제법서지이(Fasudil)대인고침습잠능HCC세포주3(human high metastatic liver cancer cells 3，HCCLM3)침습전이적영향，병차탐토기작용적궤제。방법：응용100μmol/L Fasudil작용우HCCLM3세포，채용기동단백미사형광염색화침습소실실험관찰HCCLM3세포적운동침습능력。HCCLM3세포경과처리후분위음성대조조、Fasudil작용조、BTBD7간우조，통과Western인적검측BTB/POZ결구역단백7(BR-C, ttk and bab/pox virus domain containing 7，BTBD7)、Ras동계물가족성원C(ras homolog family member C，RhoC)、Rho관련권곡라선단백격매2(Rho-associated, coiled-coil containing protein kinase 2，ROCK2)、MMP2화MMP9단백표체수평，매보분석법검측MMP2화MMP9활성수평。BTBD7간우조작위양성대조。결과：Fasudil처리후HCCLM3침습운동능력하강，BTBD7，RhoC， ROCK2단백표체하조，MMP2화MMP9활성강저，여음성대조조비교차이유통계학의의(균P<0.01)。결론：Fasudil구유간예BTBD7-ROCK2신호통로、억제HCC침습전이적중요작용。
Objective: To explore the eff ect of Fasudil on the invasion and metastatic abilities of human high metastatic liver cancer cells (HCCLM3) and the underlying mechanisms. Methods: HCCLM3 cells were incubated with 100 μmol/L Fasudil. Fluorescence staining forF-actin and Transwell assay were performed to observe the invasion ability of HCCLM3 cells. HCCLM3 cells were divided into 3 groups: a negative control group, a Fasudil group and a BTB/POZ domain containing 7 (BTBD7)-siRNA group. Western blot assay was performed to detect the expression levels of BTBD7, ras homolog family member C (RhoC) and Rho-associated, coiled-coil containing protein kinase 2 (ROCK2), matrix metalloproteinases 2 (MMP2) and MMP9. Zymogram analysis method was performed to detect the expression activities of MMP2 and MMP9. hTe BTBD7-siRNA group was served as a positive control. Results: In HCCLM3 cells treated with Fasudil, the invasion ability was significant decreased compared with the control group, concomitant with the down-regulated expression levels of BTBD7, RhoC and ROCK2 protein as well as the decreased activities of MMP2 and MMP9. Conclusion: Fasudil plays an important role in interfering BTBD7-ROCK2 signaling pathway and suppressing the invasion and metastasis of hepatocellular carcinoma.
