RhoC/ROCK-Ⅰ信号通路封闭对卵巢癌细胞生物学行为影响的研究  

Effect of the blockade of RhoC/ROCK-Ⅰ signal pathway on the biological behavior of ovarian cancer cells

作　　者：洪振亚[1] 韩志强[2] 肖敏[1] 孙立石[1] 周晓曦[1] 卢运萍[2] 周剑峰[1] 

机构地区：[1]华中科技大同济医学院附属同济医院血液科,湖北武汉430030 [2]华中科技大同济医学院附属同济医院妇产科,湖北武汉430030

出　　处：《中华肿瘤防治杂志》2009年第7期485-488,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30770914);国家重点基础研究发展规划项目(2002CB513100)

摘　　要：目的:探讨RhoC/ROCK-Ⅰ信号转导通路的封闭对卵巢癌细胞体外生物学行为的影响。方法:RT-PCR及蛋白质印迹法检测RhoC及ROCK-Ⅰ在不同卵巢癌细胞系中mR-NA及蛋白的表达水平;将ROCK-Ⅰ反义寡核苷酸(ASODN)转染人卵巢癌细胞系SW626与Caov-3细胞后,检测转染前后ROCK-Ⅰ蛋白的表达,Boyden小室观察各株细胞转染前后侵袭及运动能力的变化,MTT比色法测定转染前后黏附能力的变化。结果:RhoC和ROCK-Ⅰ在4种卵巢癌细胞中呈不同程度表达,其中RhoC的表达水平与癌细胞侵袭力和迁移能力呈正相关,r=0.95,P<0.01,转染ROCK-ⅠASODN后,细胞内ROCK-Ⅰ的表达明显减少;细胞的侵袭能力受到明显的抑制,10μmol/L组和20μmol/L组SW626细胞的侵袭能力分别为对照组的(75.6±3.8)%和(54.7±2.9)%,Caov-3为(68.8±4.7)%和(50.0±4.5)%;转染两种浓度ASODN的SW626与Caov-3细胞的随机运动能力分别为对照组的(80.0±1.3)%、(63.7±1.9)%、(72.0±1.3)%和(55.9±2.5)%;定向运动能力分别为对照组的(83.9±1.4)%、(64.1±1.3)%、(72.5±3.4)%和(54.5±1.9)%。结论:RhoC/ROCK-Ⅰ信号转导通路与人卵巢癌细胞的体外侵袭与迁移密切相关,阻断ROCK-Ⅰ的表达可有效地抑制卵巢癌肿瘤细胞的体外侵袭能力。OBJECTIVE： To investigate the effect of the activity of RhoC/ROCK-Ⅰ signal pathway on the biological behav ior of ovarian cancer cells in vitro. METHODS： The expression of RhoC and ROCK-Ⅰ in different ovarian cancer cell lines was determined by RT PCR and Western blot assay. ROCK- Ⅰ SODN was transfeeted into SW626 and Caov-3 cell lines mediated by LiofectamineTM2000. The expression of ROCK-Ⅰ after transfection was determined by the same ways above. Boyden chamber in vitro invasion assay and wound-healing assay were used to evalu- ate these cells＇ invasive and migratory capability before or after transfeetion. The changes in the adhesion of the transfected cells were detected by MTT assay. RESULTS： The expression of RhoC and ROCK- Ⅰ was different in the four cell lines, and there was a positive correlation between the level of RhoC mRNA expression and the in vitro invasive capability of all four ovarian cancer cells （r=0.95, P〈0.01）. The expressions of ROCK-Ⅰ in the cell lines were decreased significantly after transfection with ROCK- Ⅰ ASODN. When compared with control group, the invasion capability of ovarian cancer cells transfected with 10 μmol/L or 20 μmol/L ROCK-Ⅰ ASODN was inhibited to the extent of （75.6±3.8）%or （54. 7±2.9）% respectively for SW626 cell line and （68.8±4.7） % or （50.0±4.5） % for Caov3 cell line respective ly. The inhibitory effect on the chemotaxis activity of these two cell lines was （83.9±1.4）G, （64.1±1.3）G, （72.5±3.4）% and （54.5±1.9） %. CONCLUSIONS： RhoC/ROCK-Ⅰ may play a crucial role in invasion and metastasis of ovarian cancer. The in vitro invasion of ovarian cancer cell can be effectively prevented with the blockade of the RhoC/ROCK-Ⅰ signal pathway.

关 键 词：RHOC R0cK -Ⅰ 反义寡核苷酸 卵巢肿瘤/生物学 转移 

分 类 号：R737.31[医药卫生—肿瘤]