Depatment of Urology, Peking University First Hospital, Beijing, 100034, China.
Chin J Integr Med. 2023 Aug;29(8):699-706. doi: 10.1007/s11655-022-3690-9. Epub 2022 Dec 7.
To explore the effect of curcumin on the proliferation of renal cell carcinoma and analyze its regulation mechanism.
In RCC cell lines of A498 and 786-O, the effects of curcumin (2.5, 5, 10 µ mo/L) on the proliferation were analyzed by Annexin V+PI staining. Besides, A498 was inoculated into nude mice to establish tumorigenic models, and the model mice were treated with different concentrations of curcumin (100, 200, and 400 mg/kg), once daily for 30 days. Then the tumor diameter was measured, the tumor cells were observed by hematoxylin-eosin staining, and the protein expressions of miR-148 and ADAMTS18 were detected by immunohistochemistry. In vitro, after transfection of miR-148 mimics, miR-148 inhibitor or si-ADAMTS18 in cell lines, the expression of ADAMTS18 was examined by Western blotting and the cell survival rate was analyzed using MTT. Subsequently, Western blot analysis was again used to examine the autophagy phenomenon by measuring the relative expression level of LC3-II/LC3-I; autophagy-associated genes, including those of Beclin-1 and ATG5, were also examined when miR-148 was silenced in both cell lines with curcumin treatment.
Curcumin could inhibit the proliferation of RCC in cell lines and nude mice. The expression of miR-148 and ADAMTS18 was upregulated after curcumin treatment both in vitro and in vivo (P<0.05). The cell survival rate was dramatically declined upon miR-148 or ADAMTS18 upregulated. However, si-ADAMTS18 treatment or miR-148 inhibitor reversed these results, that is, both of them promoted the cell survival rate.
Curcumin can inhibit the proliferation of renal cell carcinoma by regulating the miR-148/ ADAMTS18 axis through the suppression of autophagy in vitro and in vivo. There may exist a positive feedback loop between miR-148 and ADAMTS18 gene in RCC.
探讨姜黄素对肾细胞癌增殖的影响，并分析其调控机制。
在 A498 和 786-O 肾癌细胞系中，通过 Annexin V+PI 染色分析姜黄素（2.5、5、10 μmo/L）对细胞增殖的影响。此外，将 A498 接种到裸鼠中建立肿瘤模型，并用不同浓度的姜黄素（100、200 和 400 mg/kg）处理模型小鼠，每天一次，连续 30 天。然后测量肿瘤直径，用苏木精-伊红染色观察肿瘤细胞，并通过免疫组化检测 miR-148 和 ADAMTS18 的蛋白表达。在体外，转染 miR-148 模拟物、miR-148 抑制剂或 si-ADAMTS18 后，通过 Western blot 检测 ADAMTS18 的表达，并通过 MTT 分析细胞存活率。随后，再次通过测量 LC3-II/LC3-I 的相对表达水平来通过 Western blot 分析自噬现象；当用姜黄素处理细胞系时沉默 miR-148，还检测自噬相关基因，包括 Beclin-1 和 ATG5 的表达。
姜黄素可抑制肾癌细胞系和裸鼠的增殖。姜黄素处理后，细胞和体内 miR-148 和 ADAMTS18 的表达均上调（P<0.05）。miR-148 或 ADAMTS18 上调后细胞存活率显著下降。然而，si-ADAMTS18 处理或 miR-148 抑制剂逆转了这些结果，即它们均促进了细胞存活率。
姜黄素可通过抑制自噬来抑制体外和体内肾癌细胞癌的增殖，通过调节 miR-148/ADAMTS18 轴。在肾细胞癌中，miR-148 和 ADAMTS18 基因之间可能存在正反馈回路。
