Department of Laser, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China.
Department of Dermatology, Air Force Hospital of the Western Theater Command, Chengdu, Sichuan, China.
Neoplasma. 2024 Apr;71(2):180-192. doi: 10.4149/neo_2024_240116N24.
It has been demonstrated that calreticulin (CALR) is expressed abnormally in various tumors and is involved in the occurrence and development of tumors. In this study, CALR and EIF2AK2 expression was measured in the clinical specimens of 39 patients with melanoma. Then, we constructed knockdown and overexpression cell models of CALR and EIF2AK2 and used wound healing and Transwell assays to observe cell migration and invasion. Apoptosis, EDU, and ROS assays were used to measure cell apoptosis and proliferation, as well as ROS levels. The effect of CALR on endoplasmic reticulum stress was detected using endoplasmic reticulum fluorescent probes. Western blotting was used to detect protein levels of CALR, EIF2AK2, ADAR1, and MMP14. The results indicated that CALR and EIF2AK2 expression levels were significantly higher in human melanoma tissues than in adjacent non-tumor tissue. In addition, we found a correlation between CALR and the expression of EIF2AK2 and MMP14, and the experimental results indicated that overexpression of CALR significantly upregulated the expression of EIF2AK2, MMP14, and ADAR1, while knockdown of CALR inhibited their expression. Notably, the knockdown of EIF2AK2 in the CALR overexpression group blocked the upregulation of MMP14 and ADAR1 expression by CALR, and the knockdown of both CALR and EIF2AK2 significantly inhibited MMP14 and ADAR1 expression. In conclusion, CALR and EIF2AK2 play a promoting role in melanoma progression, and knockdown of CALR and EIF2AK2 may be an effective anti-tumor target, and its mechanism may be through MMP14, ADAR1 signaling.
已经证明钙网蛋白（CALR）在各种肿瘤中异常表达，并参与肿瘤的发生和发展。在这项研究中，我们测量了 39 例黑色素瘤患者临床标本中的 CALR 和 EIF2AK2 表达。然后，我们构建了 CALR 和 EIF2AK2 的敲低和过表达细胞模型，并使用划痕愈合和 Transwell 分析观察细胞迁移和侵袭。使用凋亡、EDU 和 ROS 分析来测量细胞凋亡和增殖以及 ROS 水平。使用内质网荧光探针检测 CALR 对内质网应激的影响。使用 Western blot 检测 CALR、EIF2AK2、ADAR1 和 MMP14 的蛋白水平。结果表明，CALR 和 EIF2AK2 的表达水平在人黑色素瘤组织中明显高于相邻非肿瘤组织。此外，我们发现 CALR 与 EIF2AK2 和 MMP14 的表达之间存在相关性，实验结果表明，CALR 的过表达显著上调了 EIF2AK2、MMP14 和 ADAR1 的表达，而 CALR 的敲低抑制了它们的表达。值得注意的是，CALR 过表达组中 EIF2AK2 的敲低阻断了 CALR 对 MMP14 和 ADAR1 表达的上调，而 CALR 和 EIF2AK2 的同时敲低显著抑制了 MMP14 和 ADAR1 的表达。总之，CALR 和 EIF2AK2 在黑色素瘤进展中起促进作用，敲低 CALR 和 EIF2AK2 可能是一种有效的抗肿瘤靶点，其机制可能是通过 MMP14、ADAR1 信号通路。
