基于YAP与Akt/mTOR通路交叉对话探讨没药甾酮抑制脑胶质瘤细胞增殖的作用机制

樊昊雯; 徐宏彬

doi:10.13748/j.cnki.issn1007-7693.20241158

基于YAP与Akt/mTOR通路交叉对话探讨没药甾酮抑制脑胶质瘤细胞增殖的作用机制

Effect and Mechanism of Z-guggulsterone Inhibit the Proliferation of Glioblastoma Cells Based on the Crosstalk Between YAP and Akt/mTOR Pathway

摘要

摘要:
目的探讨没药甾酮(Z-guggulsterone，Z-GS)通过YAP与Akt/mTOR通路交叉对话调控脑胶质瘤细胞增殖的作用机制。
方法免疫组织化学染色法测定脑胶质瘤临床标本中Mst1、Lats1、YAP、p-YAP、Akt、p-Akt^Thr308、p-Akt^Ser473、mTOR、p-mTOR^Ser2448和p-mTOR^Ser2481蛋白表达；Western blotting筛选蛋白表达量较高的人脑胶质瘤细胞株。人脑胶质瘤细胞T98G经Z-GS处理后，CCK-8法检测细胞活力；平板克隆试验检测T98G细胞增殖情况；Western blotting检测T98G细胞中Mst1、p-Mst1、Lats1、p-Lats1、YAP、p-YAP、PTEN、Akt、p-Akt^Thr308、p-Akt^Ser473、mTOR、p-mTOR^Ser2448和p-mTOR^Ser2481蛋白表达。
结果与正常人脑组织相比，Ⅳ级人脑胶质瘤组织中YAP、p-YAP、Akt、p-Akt^Thr308、mTOR、p-mTOR^Ser2448阳性表达更强，p-YAP、Akt、p-mTOR^Ser2448的阳性表达与脑胶质瘤组织有显著性关联。4种人脑胶质瘤细胞中T98G细胞的上述蛋白表达量较高。Z-GS(12.5，25，50 μmol·L⁻¹)给药24 h后对人脑胶质瘤T98G细胞无细胞毒性作用且能抑制细胞增殖能力。与对照组相比，Z-GS显著下调T98G细胞中YAP、p-YAP蛋白表达和p-mTOR^Ser2448水平，显著上调p-Mst1和p-Lats1水平。
结论 Z-GS可能通过恢复Hippo通路活性，下调mTORC1活性，抑制人脑胶质瘤细胞增殖。

Abstract:
OBJECTIVE To explore the effect and mechanism of Z-guggulsterone(Z-GS) regulating glioblastoma cells proliferation through YAP and Akt/mTOR pathway crosstalk.
METHODS Determined the expression of Mst1, Lats1, YAP, p-YAP, Akt, p-Akt^Thr308, p-Akt^Ser473, mTOR, p-mTOR^Ser2448 and p-mTOR^Ser2481 proteins in glioma clinical specimens by immunohistochemical staining. Screened the glioblastoma cell lines with high expression of above proteins by Western blotting. After treatment of human glioblastoma T98G cells with Z-GS, cell viability was assessed using the CCK-8 method; and the proliferation of T98G cells was examined through plate cloning experiments; the expression of Mst1, p-Mst1, Lats1, p-Lats1, YAP, p-YAP, PTEN, Akt, p-Akt^Thr308, p-Akt^Ser473, mTOR, p-mTOR^Ser2448 and p-mTOR^Ser2481 proteins in T98G cells was determined by Western blotting.
RESULTS Compared with normal brain tissue, the positive expression of YAP, p-YAP, Akt, p-Akt^Thr308, mTOR and p-mTOR^Ser2448proteins in grade IV glioma tissue was stronger. The positive expression of p-YAP, Akt, and p-mTOR^Ser2448 was significantly correlated with the glioblastoma tissue. The expression level of above proteins in T98G cells was higher among the four glioblastoma cell lines. After T98G cells treatment with Z-GS(12.5, 25, 50 μmol·L⁻¹) for 24 h, there was no cytotoxic effects and it could inhibit the cell proliferation. Meanwhile, Z-GS significantly reduced the proteins expression of YAP, p-YAP and the level of p-mTOR^Ser2448, and significantly increased the level of p-Mst1 and p-Lats1 in T98G cells.
CONCLUSION Z-GS may inhibit the proliferation of human glioblastoma cells by restoring Hippo pathway activity and downregulating mTORC1 activity.

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