Gfi-1调节高迁移率族蛋白B1介导的自噬以克服结直肠癌中的奥沙利铂耐药性。
Gfi-1 modulates HMGB1-Mediated autophagy to overcome oxaliplatin resistance in colorectal cancer.
作者信息
Liu Weijun, Zhang Zhenyong, Zhang Liju, Jiang Xiaoming, Chen Changxian, Wu Xi, Zhao Quan
机构信息
Department of Anorectal Diseases, The First People's Hospital of Yunnan Province, Affiliated Hospital to Kunming University of Science and Technology, Kunming, 650032, PR China.
Yunnan University School Medicine, Kunming, 650032, PR China.
出版信息
Heliyon. 2024 Apr 17;10(9):e29859. doi: 10.1016/j.heliyon.2024.e29859. eCollection 2024 May 15.
BACKGROUND
Resistance to oxaliplatin (L-OHP) is a major barrier in the treatment of colorectal cancer (CRC). Autophagy is the main cause of L-OHP tolerance in CRC cells.
METHOD
The human colon cancer cell lines HCT116 and SW480 were treated with L-OHP to obtain the drug-resistant cell lines HCT116/L-OHP and SW480/L-OHP, respectively. To probe the relationship between autophagy and L-OHP tolerance of growth factor independent 1 (Gfi-1) and high-mobility group protein 1 (HMGB1) in CRC cells, gene knockout or overexpression was performed, and Western blotting was used to determine the levels of drug tolerance interrelated proteins. Transwell and CCK-8 assays were employed to analyze the proliferation of cancer cells. Immunofluorescence detection of LC3 reflected autophagy levels. Finally, the relationship between Gfi-1 and HMGB1 was detected by chromatin immunoprecipitation (ChIP).
RESULT
Compared to normal CRC cells, L-OHP-tolerant CRC cells exhibited greater autophagy (8.2 times greater in HCT116/L-OHP cells and 7.4 times greater in SW480/L-OHP cells). In addition, we detected low levels of Gfi-1 (0.6-fold for HCT116/L-OHP cells and 0.4-fold for SW480/L-OHP cells), and OE-Gfi-1 decreased HMGB1 levels (0.6-fold for HCT116/L-OHP + OE-Gfi-1 cells and 0.5-fold for SW480/L-OHP + OE-Gfi-1 cells). The inhibition of Gfi-1 further enhanced cell viability (1.7 times in HCT116+sh-Gfi-1 cells and 1.2 times in SW480+sh-Gfi-1 cells) and invasion (1.8 times in HCT116+sh-Gfi-1 cells and 2.1 times in SW480+sh-Gfi-1 cells) in CRC cells, thus promoting oxaliplatin resistance in these cells. The autophagy inhibitor 3-MA reversed the above effects. Furthermore, we noted that Gfi-1 can restrain HMGB1 expression by binding to its promoter (0.5 times in HCT116+OE-Gfi-1 cells and 0.5 times in SW480+OE-Gfi-1 cells). The inhibitory influence of 3-MA on HMGB1 reversed the influence of Gfi-1 on autophagy and malignant progression in CRC cells.
CONCLUSION
Our study suggested that Gfi-1 inhibited HMGB1 to reduce CRC autophagy levels, increasing CRC sensitivity to L-OHP.
背景
对奥沙利铂（L-OHP）耐药是结直肠癌（CRC）治疗中的主要障碍。自噬是CRC细胞对L-OHP耐受的主要原因。
方法
用L-OHP处理人结肠癌细胞系HCT116和SW480，分别获得耐药细胞系HCT116/L-OHP和SW480/L-OHP。为探究CRC细胞中自噬与生长因子独立1（Gfi-1）和高迁移率族蛋白1（HMGB1）对L-OHP耐受性之间的关系，进行基因敲除或过表达，并采用蛋白质免疫印迹法测定耐药相关蛋白水平。采用Transwell和CCK-8法分析癌细胞的增殖情况。通过免疫荧光检测LC3反映自噬水平。最后，通过染色质免疫沉淀（ChIP）检测Gfi-1与HMGB1之间的关系。
结果
与正常CRC细胞相比，耐L-OHP的CRC细胞表现出更强的自噬（HCT116/L-OHP细胞中自噬水平高8.2倍，SW480/L-OHP细胞中高7.4倍）。此外，我们检测到Gfi-1水平较低（HCT116/L-OHP细胞中为0.6倍，SW480/L-OHP细胞中为0.4倍），过表达Gfi-1可降低HMGB1水平（HCT116/L-OHP + OE-Gfi-1细胞中为0.6倍，SW480/L-OHP + OE-Gfi-1细胞中为0.5倍）。抑制Gfi-1进一步增强了CRC细胞的活力（HCT116+sh-Gfi-1细胞中为1.7倍，SW480+sh-Gfi-1细胞中为1.2倍）和侵袭能力（HCT116+sh-Gfi-1细胞中为1.8倍，SW480+sh-Gfi-1细胞中为2.1倍），从而促进这些细胞对奥沙利铂的耐药性。自噬抑制剂3-MA可逆转上述作用。此外，我们注意到Gfi-1可通过与HMGB1启动子结合来抑制其表达（HCT116+OE-Gfi-1细胞中为0.5倍，SW480+OE-Gfi-1细胞中为0.5倍）。3-MA对HMGB1的抑制作用逆转了Gfi-1对CRC细胞自噬和恶性进展的影响
结论
我们的研究表明，Gfi-1通过抑制HMGB1降低CRC自噬水平，增加CRC对L-OHP的敏感性。
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