敲低癌相关成纤维细胞中HSF1基因对口腔鳞癌细胞裸鼠皮下成瘤的影响

›› 2020, Vol. 11 ›› Issue (3): 168-172.

敲低癌相关成纤维细胞中HSF1基因对口腔鳞癌细胞裸鼠皮下成瘤的影响

姚禹¹,张嘉懿¹,刘来奎²,江宏兵³

1. 南京医科大学口腔疾病研究江苏省重点实验室
2. 南京医科大学口腔研究所
3. 南京医科大学附属口腔医院

收稿日期:2020-05-08 修回日期:2020-07-20 出版日期:2020-09-25 发布日期:2020-09-30
通讯作者: 江宏兵 E-mail:jhb@njmu.edu.cn
基金资助:
肿瘤微环境中ATP/ADO-A2aR信号轴调控巨噬细胞极化促口腔黏膜鳞癌侵袭转移机制;CAFs重塑基质微环境激活SDF1/CXCR4信号调控巨噬细胞分化与口腔癌侵袭转移机制研究

Effect of knocking down HSF1 in cancer-associated fibroblasts on subcutaneous tumor formation of oral squamous cell carcinoma in nude mice

Received:2020-05-08 Revised:2020-07-20 Online:2020-09-25 Published:2020-09-30
Contact: hongbing jiang E-mail:jhb@njmu.edu.cn

摘要/Abstract

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。

关键词: 热休克因子1, 癌相关成纤维细胞, 口腔鳞癌, 裸鼠皮下成瘤

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 CAFs-shNC 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.

Key words: HSF1, CAFs, oral squamous cell carcinoma, subcutaneous tumor formation in nude mice.

姚禹张嘉懿刘来奎江宏兵. 敲低癌相关成纤维细胞中HSF1基因对口腔鳞癌细胞裸鼠皮下成瘤的影响[J]. 口腔生物医学, 2020, 11(3): 168-172.

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