摘要
目的探讨热疗调控TPX2表达诱导胆管癌细胞的凋亡机制。方法在不同温度(37℃、40℃、43℃和46℃)下处理后,检测胆管癌细胞RBE的凋亡水平和细胞活力。热疗处理后,转录组高通量测序和siRNA筛选鉴定热疗调控胆管癌细胞RBE凋亡的关键分子。结果随着温度升高(37℃、40℃、43℃和46℃),胆管癌细胞的活力降低(P<0.05),坏死的胆管癌细胞随着温度的升高而增加(P<0.05),细胞凋亡水平在43℃时达到峰值(6%±1%,P<0.05)。当敲低TPX2时,胆管癌细胞的凋亡水平上升(47%±3%比74%±3%,P<0.05)。过表达TPX2时,胆管癌细胞的凋亡水平下降(45%±3%比21%±4%,P<0.05)。热疗显著降低了TPX2、PI3K、p-PI3K和P-AKT的表达水平(P<0.05),并增加了p21的表达和caspase3的切割(P<0.05);敲低TPX2后,PI3K、p-PI3K和p-AKT的表达水平显著下降(P<0.05),p21的表达水平显著上升(P<0.05);过表达TPX2后,PI3K和p-AKT的表达水平显著上升(P<0.05),p21的表达水平显著下降(P<0.05)。结论热疗通过影响TPX2的表达,抑制了PI3K/AKT通路活性,减少了胆管癌细胞的细胞活力,促进了胆管癌细胞的凋亡。
Objective To investigate the mechanism underlying apoptosis of cholangiocarcinoma(CCA)cells induced by hyperthermia-regulated intracellular expression of TPX2.Methods The apoptosis and cell viability of a CCA cell-line RBE were measured after treatment under different temperatures(37℃,40℃,43℃and 46℃).Following hyperthermia,high-throughput sequencing of transcriptome and siRNA screening were performed to identify the key molecules that underpin the hyperthermia regulation of CCA RBE cell apoptosis.Results The viability of CCA cells declined with ascending temperatures(37℃,40℃,43℃and 46℃)(P<0.05),paralleled with increasing number of necrotic CCA cells(P<0.05).The cell apoptosis level peaked at 43℃(6%±1%,P<0.05).Knockout of TPX2 led to a higher level of CCA cell apoptosis(47%±3%vs 74%±3%,P<0.05);conversely,the CCA cell apoptosis decreased with TPX2 overexpression(45%±3%vs 21%±4%,P<0.05).Hyperthermia was significantly linked to lowered expression of TPX2,PI3K,p-PI3K and P-AKT(P<0.05),and elevated p21 expression and caspase3 cleavage(P<0.05).After TPX2 knockout,significantly there were reductions in the expression of PI3K,p-PI3K and p-AKT(P<0.05)and an increase in p21 expression(P<0.05).After overexpression of TPX2,the expression levels of PI3K and p-AKT were significantly increased(P<0.05)and that of p21 was significantly decreased(P<0.05).Conclusion Hyperthermia suppresses activation of the PI3K/AKT pathway via regulating the TPX2 expression,thereby attenuates viability and promotes apoptosis of CCA cells.
作者
贺艳平
李江怀
He Yanping;Li Jianghuai(Department of Hepatobiliary and Pancreatic Surgery,Shanxi Provincial People’s Hospital,Taiyuan 030012,China)
出处
《中华生物医学工程杂志》
CAS
2022年第6期623-629,共7页
Chinese Journal of Biomedical Engineering
关键词
高温
诱发
胆管肿瘤
细胞凋亡
信号通路
Hyperthermia,induced
Bile duct neoplasms
Apoptosis
Signaling pathway
