Gu Yong-hong, Zhou Chang-ju, Hu Lin-yu, Chen Qian, Zhang Wei-she
Department of Pathology, Third Xiangya Hospital, Central South University, Changsha 410013, China. guyonghong
Nan Fang Yi Ke Da Xue Xue Bao. 2010 Jun;30(6):1369-72.
To study the effects of calponin-1 expression inhibition on the proliferation , invasiveness, apoptosis and cytoskeleton of uterine smooth muscle cells, and explore the molecular mechanism of calponin-1 in the uterine smooth muscle cells for labor onset.
siRNA-calponin-1 adenovirus plasmid was constructed and transfected into primarily cultured uterine smooth muscle cells. The proliferation, invasiveness and apoptosis of the cells were determined by MTT assay, matrigel invasion assays and flow cytometry, respectively. Rhodamine-Phalloidin was used for labeling filamentous actin (F-actin), and the morphology and the distribution of F-actin was observed under fluorescence microscopy and analyzed quantitatively.
The motor ability of uterine smooth muscle cells decreased significantly after transfection with siRNA-calponin-1 adenovirus plasmid (P<0.05). The transfected cells showed thinner, loosened and irregular F-actin microfibers, and the cells in the empty vector and blank control groups showed thicker and longer F-actin microfibers.
Inhibition of calponin-1 expression can inhibit uterine smooth muscle cell migration and cause the morphological change and rearrangement of F-actin without affecting its proliferation and apoptosis in vitro, suggesting that the morphological change and rearrangement of F-actin of uterine smooth muscle cell may be one of the important mechanisms in the labor onset.
研究钙调蛋白-1表达抑制对子宫平滑肌细胞增殖、侵袭、凋亡及细胞骨架的影响,探讨钙调蛋白-1在子宫平滑肌细胞发动分娩中的分子机制。
构建siRNA-钙调蛋白-1腺病毒质粒并转染至原代培养的子宫平滑肌细胞。分别采用MTT法、基质胶侵袭实验和流式细胞术检测细胞的增殖、侵袭及凋亡情况。用罗丹明-鬼笔环肽标记丝状肌动蛋白(F-肌动蛋白),在荧光显微镜下观察F-肌动蛋白的形态和分布并进行定量分析。
转染siRNA-钙调蛋白-1腺病毒质粒后子宫平滑肌细胞的运动能力显著降低(P<0.05)。转染后的细胞F-肌动蛋白微丝更细、更松散且不规则,空载体组和空白对照组的细胞F-肌动蛋白微丝更粗、更长。
抑制钙调蛋白-1表达可抑制子宫平滑肌细胞迁移并导致F-肌动蛋白的形态改变和重排,而在体外不影响其增殖和凋亡,提示子宫平滑肌细胞F-肌动蛋白的形态改变和重排可能是发动分娩的重要机制之一。