Chen Shaomu, Huang Haitao, Yao Jie, Pan Liangbin, Ma Haitao
Department of Cardiothoracic Surgery, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China.
Mol Med Rep. 2014 Aug;10(2):677-82. doi: 10.3892/mmr.2014.2240. Epub 2014 May 15.
The aim of the present study was to address the effects of heat shock protein B6 (HspB6) on tumor growth and metastasis in BALB/c mice. Lewis lung carcinoma (LLC) cells were subcutaneously injected into BALB/c mice followed by intraperitoneal injection of recombinant HspB6 (HspB6 groups) or phosphate‑buffered saline (control groups). Tumor growth and metastasis were assessed by size measurement and weighing of tumors and cervical lymph nodes, respectively. Chemokine expression in tumor masses was quantified quantitative polymerase chain reaction and western blotting. Tumor cell apoptosis was detected by flow cytometric analysis. The proliferation and migration of LLC cells, stimulated with HspB6, were detected using Cell Counting Kit 8 and wound scratch assays in vitro. Tumors grafted into the BALB/c mice and intraperitoneally injected with HspB6 were significantly bigger in size than those grafted into the control mice. From 7 days following the injection, the weight of cervical lymph nodes in HspB6 groups was higher than that in the control mice. We also revealed that the apoptotic cell number in tumor masses in the HspB6 groups was lower than that of the control mice. CD31 expression of vascular endothelial cells was higher in tumors grafted in HspB6 groups than those grafted in the control mice. Concomitantly, the tumor tissue mRNA and protein expression enhancement of vascular endothelial growth factor, basic fibroblast growth factor and intercellular adhesion molecule 1 were greater in HspB6 mice than in the control mice. HspB6 also inhibited cell apoptosis and enhanced the migration and proliferation of LLCs in vitro. In conclusion, HspB6 exhibited tumor promotion through increasing tumor angiogenesis, tumor metastasis and inhibiting tumor cell apoptosis.
本研究的目的是探讨热休克蛋白B6（HspB6）对BALB/c小鼠肿瘤生长和转移的影响。将Lewis肺癌（LLC）细胞皮下注射到BALB/c小鼠体内，随后腹腔注射重组HspB6（HspB6组）或磷酸盐缓冲盐水（对照组）。分别通过测量肿瘤和颈部淋巴结的大小及称重来评估肿瘤生长和转移情况。通过定量聚合酶链反应和蛋白质印迹法对肿瘤块中的趋化因子表达进行定量分析。采用流式细胞术分析检测肿瘤细胞凋亡情况。在体外，使用细胞计数试剂盒8和划痕实验检测经HspB6刺激的LLC细胞的增殖和迁移情况。移植到BALB/c小鼠体内并腹腔注射HspB6的肿瘤，其大小明显大于移植到对照小鼠体内的肿瘤。注射后7天起，HspB6组颈部淋巴结的重量高于对照小鼠。我们还发现，HspB6组肿瘤块中的凋亡细胞数量低于对照小鼠。HspB6组移植肿瘤中血管内皮细胞的CD31表达高于对照小鼠移植肿瘤。同时，HspB6小鼠肿瘤组织中血管内皮生长因子、碱性成纤维细胞生长因子和细胞间黏附分子1的mRNA和蛋白表达增强程度大于对照小鼠。HspB6在体外也抑制细胞凋亡并增强LLC细胞的迁移和增殖。总之，HspB6通过增加肿瘤血管生成、促进肿瘤转移和抑制肿瘤细胞凋亡发挥促肿瘤作用。
