Page 42 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第42卷第5期
·638 · Journal of Nanjing Medical University（Natural Sciences） 2022年5月

·基础研究·

缺氧通过HIF⁃1α介导细胞内YAP1降低骨肉瘤细胞对CDDP的

敏感性及机制

黄煜龙，李晨露
东南大学附属中大医院药学部，江苏 南京 210009

［摘 要］ 目的：研究 Yes 相关蛋白 1（yes⁃associated protein 1，YAP1）对缺氧条件下骨肉瘤细胞顺铂（cisplatin，CDDP）敏感性
的影响及潜在机制。方法：采用流式细胞术和CCK⁃8检测骨肉瘤MG⁃63细胞对CDDP的敏感性。Western blot方法检测MG⁃63
细胞在常氧和缺氧条件下、转染HIF⁃1α⁃siRNA、YAP1⁃siRNA、STAT3⁃siRNA后，细胞内缺氧诱导因子1α（hypoxia inducible fac⁃
tor⁃1α，HIF⁃1α）、YAP1、信号转导和转录激活因子3（signal transducer and activator of transcription 3，STAT3）和磷酸化STAT3表
达水平。结果：YAP1在缺氧后的人骨肉瘤细胞MG⁃63中呈显著高表达（P < 0.05）。缺氧条件下过表达YAP1，MG⁃63细胞对
CDDP 的敏感性显著下降（P < 0.01）。缺氧条件下采用 siRNA 抑制 YAP1 表达，MG⁃63 细胞对 CDDP 的敏感性显著提高（P <
0.01）；细胞中 p⁃STAT3 表达显著下调（P < 0.01）。沉默 STAT3 可显著提高缺氧条件下 MG⁃63 细胞对 CDDP 的敏感性（P <
0.01）。沉默 HIF⁃1α可显著抑制缺氧诱导的 YAP1 表达上调（P < 0.01），提高缺氧条件下 MG⁃63 细胞对 CDDP 的敏感性（P <
0.01）。结论：缺氧通过HIF⁃1α依赖的方式上调骨肉瘤细胞内YAP1，YAP1可能通过激活STAT3降低骨肉瘤细胞对CDDP的敏
感性。
［关键词］ 骨肉瘤；YAP1；STAT3；缺氧；耐药
［中图分类号］ R738.1 ［文献标志码］ A ［文章编号］ 1007⁃4368（2022）05⁃638⁃06
doi：10.7655/NYDXBNS20220506

Hypoxia decreases the sensitivity of osteosarcoma cells to CDDP by regulating HIF⁃1α⁃
mediated YAP1 and its mechanism

HUANG Yulong，LI Chenlu
Department of Pharmacy，Zhongda Hospital Affiliated to Southeast University，Nanjing 21009，China

［Abstract］ Objective：This study aims to investigate the effect of yes associated protein 1（YAP1）on cisplatin（CDDP）sesitivity of
osteosarcoma cells under hypoxia and its underlying mechanism. Methods：Flow cytometry and CCK⁃8 were used to detect the CDDP
sensitivity of osteosarcoma cells. Western blot was used to detect the expression of hypoxia inducible factor⁃1α（HIF⁃1α），YAP1，signal
transducer and activator of transcription 3（STAT3）and p⁃STAT3 protein in human osteosarcoma cell line MG⁃63 under normoxic and
hypoxic conditions after transfection with HIF⁃1 α⁃siRNA，YAP1⁃siRNA and STAT3⁃siRNA. Results：YAP1 was highly expressed in
MG⁃63 cells after hypoxia（P < 0.05）. Overexpression of YAP1 significantly reduced the sensitivity of MG⁃63 cells to CDDP under
hypoxic condition（P < 0.01）. The apoptosis rate of osteosarcoma cells was significantly increased（P < 0.01）and the sensitivity of
osteosarcoma cells to CDDP was increased（P < 0.01）under hypoxic condition after transfection with YAP1⁃siRNA. The expression of p
⁃STAT3 in MG⁃63 cells was significantly down⁃regulated after inhibiting YAP1 expression（P < 0.01）. Silencing STAT3 significantly
increased the sensitivity of MG⁃63 cells to CDDP under hypoxia（P < 0.01）. Silencing HIF⁃1α significantly inhibited hypoxia⁃induced
up⁃regulation of YAP1（P < 0.01），and incresed the sensitivity of MG⁃63 cells to CDDP. Conclusion：Hypoxia up⁃regulates YAP1 in
osteosarcoma cells in a HIF⁃1α dependent manner. YAP1 may promote CDDP resistance of osteosarcoma cells by activating STAT3.
［Key words］ osteosarcoma；YAP1；STAT3；hypoxia；drug resistance
［J Nanjing Med Univ，2022，42（05）：638⁃643］