Li Ruohan, Huang Yingzhao, Liao Nailin, Wu Chenzhou, Li Yi
State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Head and Neck Oncology Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2022 Jan 25;40(1):22-31. doi: 10.7518/hxkq.2022.01.004.
In this study, we aimed to investigate whether oral cancer cells affect pancreatic β-cells function through transmissible endoplasmic reticulum stress (TERS).
Tunicamycin (TM) was selected as the endoplasmic reticulum stress (ERS) inducer. The human oral cancer cell lines CAl-27 and SCC-25 were selected as the donor cells, and mouse insulinoma 6 (MIN6) cell lines were chosen as the recipient cells. Quantitative real-time polymerase chain reaction (qPCR) and Western blot (WB) analysis were used to detect ERS markers and insulin expression. The TdT-mediated dUTP nick-end labeling (TUNEL) method was applied to detect apoptosis levels. The clone formation method was utilized to detect cell proliferation capability. The secretory function of pancreatic β-cells was detected with an enzyme linked immunosorbent assay (ELISA) kit and a bicinchoninic acid (BCA) kit.
The MIN6 cells were subjected to TM stimulation. qPCR and WB analysis revealed that ERS markers were upregulated. This result implied that the MIN6 cells can induce ERS. The supernatant of oral cancer cells under ERS was added to the MIN6 cells. qPCR and WB analysis showed that the oral cancer cells that had been subjected to ERS could induce ERS in the MIN6 cells, that is, the phenomenon of TERS occurred. The TUNEL assay indicated that the apoptosis of the MIN6 cells increased under TERS. The clone formation assay demonstrated that the proliferation capability of the MIN6 cells decreased under TERS. qPCR and WB analysis revealed that under TERS, insulin synthesis by the MIN6 cells decreased and insulin synthesis was inhibited at the translation level. The ELISA and BCA kits demonstrated that insulin secretion by the MIN6 cells was reduced under TERS.
Oral cancer cells can affect pancreatic β-cells through TERS, resulting in increased apoptosis, decreased viability, and reduced insulin secretion and synthesis capability.
在本研究中，我们旨在调查口腔癌细胞是否通过可传播的内质网应激（TERS）影响胰腺β细胞功能。
选择衣霉素（TM）作为内质网应激（ERS）诱导剂。选择人类口腔癌细胞系CAl-27和SCC-25作为供体细胞，选择小鼠胰岛素瘤6（MIN6）细胞系作为受体细胞。采用定量实时聚合酶链反应（qPCR）和蛋白质免疫印迹（WB）分析来检测ERS标志物和胰岛素表达。应用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）法检测凋亡水平。利用克隆形成法检测细胞增殖能力。用酶联免疫吸附测定（ELISA）试剂盒和二喹啉甲酸（BCA）试剂盒检测胰腺β细胞的分泌功能。
对MIN6细胞进行TM刺激。qPCR和WB分析显示ERS标志物上调。该结果表明MIN6细胞可诱导ERS。将ERS状态下口腔癌细胞的上清液添加到MIN6细胞中。qPCR和WB分析表明，经历ERS的口腔癌细胞可诱导MIN6细胞发生ERS，即发生了TERS现象。TUNEL检测表明，在TERS作用下MIN6细胞的凋亡增加。克隆形成试验表明，在TERS作用下MIN6细胞的增殖能力下降。qPCR和WB分析显示，在TERS作用下，MIN6细胞的胰岛素合成减少，且胰岛素合成在翻译水平受到抑制。ELISA和BCA试剂盒表明，在TERS作用下MIN6细胞的胰岛素分泌减少。
口腔癌细胞可通过TERS影响胰腺β细胞，导致凋亡增加、活力降低以及胰岛素分泌和合成能力下降。
