KW-2478对结直肠癌细胞增殖的抑制作用及其机制

doi:10.3969/j.issn.1004-616x.2024.03.006

癌变·畸变·突变 ›› 2024, Vol. 36 ›› Issue (3): 202-207.doi: 10.3969/j.issn.1004-616x.2024.03.006

KW-2478对结直肠癌细胞增殖的抑制作用及其机制

肖雪, 卢晓童, 陈思琦, 姜玉娟, 郝佳洁, 蔡岩, 王明荣, 张钰

国家癌症中心/国家肿瘤临床医学研究中心/中国医学科学院北京协和医学院肿瘤医院, 分子肿瘤学国家重点实验室, 北京 100021

收稿日期:2024-03-13 修回日期:2024-04-29 出版日期:2024-05-30 发布日期:2024-06-05
通讯作者: 张钰
作者简介:肖雪，E-mail：bioxiaoxue@foxmail.com。
基金资助:
国家自然科学基金(82073093)

Inhibitory effect and underlying mechanism of KW-2478 on proliferation of colorectal cancer cells in vitro

XIAO Xue, LU Xiaotong, CHEN Siqi, JIANG Yujuan, HAO Jiajie, CAI Yan, WANG Mingrong, ZHANG Yu

State Key Laboratory of Molecular Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China

Received:2024-03-13 Revised:2024-04-29 Online:2024-05-30 Published:2024-06-05

摘要/Abstract

摘要： 目的：探讨HSP90α抑制剂KW-2478对结直肠癌细胞恶性表型的影响并研究其作用机制。方法：以溶剂DMSO为对照，采用不同浓度的HSP90α小分子抑制剂KW-2478处理结直肠癌RKO细胞和DLD1细胞。采用CCK8试剂盒检测结直肠癌细胞的增殖率；平板集落形成实验检测细胞集落形成率；流式细胞术分析细胞周期；Western blot法检测细胞中增殖和周期调控相关蛋白的表达水平及磷酸化水平。结果：与DMSO溶剂对照组比较，0.8 μmol/L KW-2478处理后的RKO细胞与40 μmol/L KW-2478处理后的DLD1细胞增殖率均降低50%以上；KW-2478对RKO细胞和DLD1细胞的IC₅₀分别为(0.5±1.2) μmol/L和(40.0±3.1) μmol/L。KW-2478处理后，RKO细胞和DLD1细胞的集落形成率降低40%以上(均为P<0.01)。同时，KW-2478可显著诱导RKO细胞和DLD1细胞发生G2/M期阻滞(均为P<0.01)。与DMSO溶剂对照组相比，KW-2478处理的RKO和DLD1细胞中HSP90α客户蛋白EGFR和AKT、S6，以及p-AKT、p-ERK和p-S6蛋白的表达水平均降低。同时，相较于对照组，KW-2478处理组细胞中G2/M期分子标志p-Histone H3以及Cyclin B1蛋白的表达水平升高。结论：KW-2478可诱导结直肠癌细胞系RKO和DLD1发生明显的G2/M期阻滞并显著抑制细胞的增殖，该作用可能与其抑制EGFR相关信号通路活性及上调周期相关蛋白的表达有关。

关键词: 结直肠癌, 热休克蛋白90, KW-2478, 增殖能力, 细胞周期, 表皮生长因子受体

Abstract: OBJECTIVE：To investigate effect and underlying mechanism of HSP90α inhibitor KW-2478 on malignant phenotypes of colorectal cancer (CRC) cells. METHODS：Using DMSO as solvent control，colorectal cancer cells RKO and DLD1 were treated with different concentrations of KW-2478. The proliferation rate of colorectal cancer cells was detected by CCK8 kit；the colony formation rate was detected by plate colony formation assay；flow cytometry was used to analyze the cell cycle；Western blot was used to detect the expression level and phosphorylation level of proliferation and cycle regulation-related proteins. RESULTS：Compared with the control group，the proliferation capacity of RKO cells treated with 0.8 μmol/L KW-2478 and DLD1 cells treated with 40 μmol/L KW-2478 decreased by more than 50%，and the IC₅₀ of both cells were (0.5±1.2) μmol/L and (40.0±3.1) μmol/L，respectively. After treatment with KW-2478，the sum colony formation rate of both cells were reduced by more than 40% when compared with the control group (P<0.01) . Meanwhile，KW-2478 significantly induced G2/M phase arrest (P<0.01). Upon KW-2478 treatment，the protein abundance of HSP90a client proteins EGFR，AKT and S6，and the phosphorylation levels of AKT，ERK and S6 were reduced，while the expressions of mitosis-specific marker p-Histone H3 and Cyclin B1 protein were upregulated. CONCLUSION：KW-2478 significantly inhibited the proliferation viability and colony formation ability of RKO and DLD1 and induced markedly G2/M phase arrest. The observed effects may be related to inhibiting the activity of EGFR-related signaling pathways and upregulating the expression of cycle-related proteins.

Key words: colorectal cancer, HSP90, KW-2478, proliferation, cell cycle, EGFR

中图分类号:

R735.1

肖雪, 卢晓童, 陈思琦, 姜玉娟, 郝佳洁, 蔡岩, 王明荣, 张钰. KW-2478对结直肠癌细胞增殖的抑制作用及其机制[J]. 癌变·畸变·突变, 2024, 36(3): 202-207.

XIAO Xue, LU Xiaotong, CHEN Siqi, JIANG Yujuan, HAO Jiajie, CAI Yan, WANG Mingrong, ZHANG Yu. Inhibitory effect and underlying mechanism of KW-2478 on proliferation of colorectal cancer cells in vitro[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(3): 202-207.

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KW-2478对结直肠癌细胞增殖的抑制作用及其机制

Inhibitory effect and underlying mechanism of KW-2478 on proliferation of colorectal cancer cells in vitro

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