Lin Chunlong, Li Xiaohui, Luo Qiong, Yang Hui, Li Lun, Zhou Qiong, Li Yue, Tang Hao, Wu Lifu
Respiratory Department of the Second Hospital of Yueyang, Hunan Normal University, Yueyang, Hunan 414000, China.
Respiratory Department of the Second Hospital of Yueyang, Hunan Normal University, Yueyang, Hunan 414000, China.
Exp Cell Res. 2017 Feb 1;351(1):43-50. doi: 10.1016/j.yexcr.2016.12.021. Epub 2016 Dec 30.
Resistin-like molecule-β (RELM-β), focal adhesion kinase (FAK), and survivin may be involved in the proliferation of cultured human pulmonary artery smooth muscle cells (HPAMSCs), which is involved in pulmonary hypertension. HPAMSCs were treated with human recombinant RELM-β (rhRELM-β). siRNAs against FAK and survivin were transfected into cultured HPASMCs. Expression of FAK and survivin were examined by RT-PCR and western blot. Immunofluorescence was used to localize FAK. Flow cytometry was used to examine cell cycle distribution and cell death. Compared to the control group, all rhRELM-β-treated groups demonstrated significant increases in the expression of FAK and survivin (P<0.05). rhRELM-β significantly increased the proportion of HPASMCs in the S phase and decreased the proportion in G0/G1. FAK siRNA down-regulated survivin expression while survivin siRNA did not affect FAK expression. FAK siRNA effectively inhibited FAK and survivin expression in RELM-β-treated HPASMCs and partially suppressed cell proliferation. RELM-β promoted HPASMC proliferation and upregulated FAK and survivin expression. In conclusion, results suggested that FAK is upstream of survivin in the signaling pathway mediating cell proliferation. FAK seems to be important in RELM-β-induced HPASMC proliferation, partially by upregulating survivin expression.
抵抗素样分子β（RELM-β）、粘着斑激酶（FAK）和生存素可能参与了培养的人肺动脉平滑肌细胞（HPAMSCs）的增殖，而这一过程与肺动脉高压有关。用重组人RELM-β（rhRELM-β）处理HPAMSCs。将针对FAK和生存素的小干扰RNA（siRNAs）转染到培养的HPASMCs中。通过逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测FAK和生存素的表达。采用免疫荧光法对FAK进行定位。运用流式细胞术检测细胞周期分布和细胞死亡情况。与对照组相比，所有rhRELM-β处理组的FAK和生存素表达均显著增加（P<0.05）。rhRELM-β显著增加了处于S期的HPASMCs比例，降低了处于G0/G1期的比例。FAK小干扰RNA下调了生存素的表达，而生存素小干扰RNA不影响FAK的表达。FAK小干扰RNA有效抑制了rhRELM-β处理的HPASMCs中FAK和生存素的表达，并部分抑制了细胞增殖。RELM-β促进了HPASMC的增殖，并上调了FAK和生存素的表达。总之，结果表明在介导细胞增殖的信号通路中，FAK在生存素的上游。FAK似乎在RELM-β诱导的HPASMC增殖中起重要作用，部分是通过上调生存素的表达来实现的。
