【摘要】 目的:探讨癌相关成纤维细胞(CAFs)中热休克因子1(HSF1)基因对口腔鳞癌(OSCC)细胞裸鼠皮下移植瘤生长和转移的影响。方法:采用Western blot鉴定体外分离培养的OSCC组织及癌旁组织成纤维细胞,以获取CAFs;采用慢病毒敲低CAFs中的HSF1,构建CAFs-shHSF1细胞(实验组)和CAFs-shNC细胞(对照组);将肿瘤细胞Cal27分别与CAFs-shHSF1细胞和CAFs-shNC细胞按1∶1混匀,接种于裸鼠腋下,观察成瘤情况及移植瘤生长变化,分析体质量和瘤体体积差异,HE染色和免疫组化染色观察移植瘤转移以及上皮间质转化(EMT)情况。结果:OSCC组织成纤维细胞中α-平滑肌肌动蛋白(α-SMA)、成纤维细胞活化蛋白(FAP)和成纤维细胞特异蛋白-1(FSP-1)等CAFs标记物的蛋白丰度较癌旁组织成纤维细胞明显升高。实验组裸鼠的肿瘤体积明显低于对照组(P<0.05),而裸鼠体质量没有显著差异(P>0.05);实验组肝转移瘤数目少于对照组(P<0.05);与对照组比较,实验组移植瘤α-SMA和波形蛋白免疫组化染色减弱,上皮细胞钙粘蛋白染色无明显改变。结论:敲低CAFs中的HSF1基因能够显著抑制OSCC细胞裸鼠移植瘤的生长和转移,可能通过部分EMT机制发挥作用。更多还原

【Abstract】 Objective: To investigate the effect of HSF1 in cancer-associated fibroblast( CAFs) on the growth and metastasis of subcutaneous xenotransplanted tumor of oral squamous cell carcinoma( OSCC) cell in nude mice. Methods: Fibroblasts of OSCC and adjacent normal tissues were identified by Western blot to obtain CAFs. CAFs-shHSF1 cells and control CAFs-shNC cells were constructed by lentivirus knockdown HSF1 in CAFs. The tumor cells Cal27 were mixed respectively with CAFs-shHSF1 cells and CAFssh NC cells at 1 ∶ 1 and inoculated into the right axilla of nude mice,and the formation and growth of the xenografted tumor were observed. The difference of tumor volume and weight between the two groups was statistically analyzed. HE staining and immunohistochemical staining were used to verify the metastasis and epithelial-mesenchymal transformation( EMT) of the xenografted tumor between the two groups. Results: The protein abundance of α-SMA,FAP and FSP-1,the markers of CAFs,was significantly higher than those of normal fibroblasts. The tumor volume in the experimental group was significantly lower than that in the control group( P<0.05),and there was no significant difference in body weight between the two groups( P>0.05). Meanwhile,there was liver metastasis in both the experimental group and the control group,and the number of liver metastasis in the experimental group was less than that in the control group( P<0.05). Immunohistochemical staining data showed that α-SMA and Vimentin staining decreased and E-cadherin did not change significantly in Cal27 +CAFs-shHSF1 group. Conclusions: Knockdown of HSF1 in CAFs can significantly inhibit the growth and metastasis of OSCC in nude mice,which may play a role through partial EMT mechanism.更多还原