大黄素通过调节自噬抑制口腔鳞状细胞癌细胞的增殖

doi:10.13591/j.cnki.kqyx.2024.08.009

摘要/Abstract

摘要：

目的探究大黄素对口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)细胞增殖的影响及其作用机制。方法用不同浓度的大黄素干预CAL-27细胞、SCC-15细胞,CCK-8法检测细胞的增殖活性,并筛选出合适的浓度进行后续实验,集落形成实验检测大黄素对OSCC细胞集落形成能力的影响。蛋白免疫印迹法(Western blot)检测增殖相关蛋白PCNA,自噬相关蛋白Beclin1、LC3的表达;联合雷帕霉素与一定浓度的大黄素共处理细胞后,CCK-8检测细胞增殖活力,集落形成实验检测细胞集落形成能力,Western blot检测PCNA、Beclin1、LC3的表达。裸鼠成瘤实验检测大黄素对裸鼠瘤体瘤重、瘤体积的影响,HE染色观察裸鼠肝脏及肾脏结构,IHC检测PCNA、Beclin1蛋白的表达情况。结果大黄素可显著降低CAL-27细胞的增殖活性及集落形成能力,且呈现浓度依赖性,大黄素能下调PCNA、Beclin1、LC3Ⅱ/LC3Ⅰ的表达量,RAPA可逆转大黄素对细胞的增殖抑制及对增殖、自噬等蛋白表达的影响。体内实验显示,大黄素无明显的肝、肾毒性,可降低瘤体的体积和质量,下调瘤体中PCNA、Beclin1的蛋白表达。结论大黄素可抑制口腔鳞癌细胞的增殖能力,其作用可能与抑制细胞自噬相关。

关键词: 大黄素, 自噬, 口腔鳞癌, 增殖, 雷帕霉素

Abstract:

Objective To investigate the effect and mechanism of emodin on cell proliferation in OSCC. Methods CAL-27 cells and SCC-15 cells were intervened with different concentrations of emodin, and the proliferative activity of the cells was detected by CCK-8 method, and the appropriate concentration was screened for follow-up experiments. The colony formation experiment helped detect the colony formation ability. The expressions of PCNA, Beclin1 and LC3 were detected using Western blot. After treating the cells with rapamycin and emodin, the cell proliferation activity and the colony formation ability were examined. PCNA, Beclin1 and LC3 expressions were detected. The tumor formation experiment helped detect the effect of emodin on tumor weight and volume in nude mice. The liver and kidney structure in nude mice was observed using HE staining. Moreover, the PCNA and Beclin1 protein expression was detected by IHC. Results Emodin could significantly decrease the proliferative activity and aggregation ability of CAL-27 cells in a concentration-dependent manner. Emodin could down-regulate the PCNA, Beclin1, and LC3Ⅱ/LC3Ⅰ expression levels. RAPA could reverse the inhibitory effect of emodin on cell proliferation. Emodin had no significant hepatorenal toxicity and could reduce the volume and mass of the tumor and down-regulate the expression of PCNA and Beclin1 protein inside the tumor. Moreover, the expression of PCNA and Beclin1 in tumor showed a certain positive correlation. Conclusion Emodin can inhibit oral squamous cell proliferation, and its effect may be associated with autophagy inhibition.

Key words: emodin, autophagy, oral squamous cell carcinoma, proliferation, rapamycin

中图分类号:

R739.85

熊轲, 张昊, 胡图强. 大黄素通过调节自噬抑制口腔鳞状细胞癌细胞的增殖[J]. 口腔医学, 2024, 44(8): 602-608.

XIONG Ke, ZHANG Hao, HU Tuqiang. Experimental study on emodin regulating the proliferation of oral squamous cell carcinoma cells through autophagy[J]. Stomatology, 2024, 44(8): 602-608.

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