c-Met抑制剂AMG-102增强喉鳞癌细胞放射敏感性的机制
目的 探讨c?Met抑制剂AMG?102对喉鳞癌细胞的增殖抑制和放射增敏作用.方法 采用四甲基偶氮唑蓝(MTT)法检测AMG?102对喉鳞癌细胞株Hep?2和KBV200的增殖抑制作用.采用克隆形成实验分析AMG?102对Hep?2和KBV200细胞的放射增敏作用.采用流式细胞术检测Hep?2和KBV200细胞的凋亡情况.采用Western blot法检测细胞中c?Met／p?Met、凋亡蛋白cleaved caspase 3及下游信号通路蛋白Akt／p?Akt和Erk／p?Erk的表达.利用RNA干扰技术下调细胞中c?Met表达,转染c?Met过表达质粒使细胞中c?Met过表达,分析Hep?...
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| Published in | 中华肿瘤杂志 Vol. 41; no. 12; pp. 909 - 917 |
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| Main Authors | , , , , , , , , |
| Format | Journal Article |
| Language | Chinese |
| Published |
河北医科大学第四医院放疗科,石家庄,050011%邢台市柏乡县中心医院医务科055450%河北医科大学第一医院耳鼻喉科,石家庄,050031%解放军联勤保障部队第九八〇医院放疗科,石家庄,050082%解放军联勤保障部队第九八〇医院耳鼻喉头颈外科,石家庄,050082%解放军联勤保障部队第九八〇医院肿瘤科,石家庄,050082
01.12.2019
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| Online Access | Get full text |
| ISSN | 0253-3766 |
| DOI | 10.3760/cma.j.issn.0253-3766.2019.12.006 |
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| Abstract | 目的 探讨c?Met抑制剂AMG?102对喉鳞癌细胞的增殖抑制和放射增敏作用.方法 采用四甲基偶氮唑蓝(MTT)法检测AMG?102对喉鳞癌细胞株Hep?2和KBV200的增殖抑制作用.采用克隆形成实验分析AMG?102对Hep?2和KBV200细胞的放射增敏作用.采用流式细胞术检测Hep?2和KBV200细胞的凋亡情况.采用Western blot法检测细胞中c?Met／p?Met、凋亡蛋白cleaved caspase 3及下游信号通路蛋白Akt／p?Akt和Erk／p?Erk的表达.利用RNA干扰技术下调细胞中c?Met表达,转染c?Met过表达质粒使细胞中c?Met过表达,分析Hep?2和KBV200细胞对AMG?102的敏感性变化.结果 与KBV200细胞比较,Hep?2细胞对AMG?102更加敏感,半数抑制浓度(IC50)分别为14和9 μmol／L. Hep?2细胞中c?Met和p?Met蛋白的相对表达量分别为194.48±0.57和177.76±1.53,均明显高于KBV200细胞(分别为171.24±1.00和115.37±0.56,均P＜0.001). AMG?102作用于肝细胞生长因子(HGF)处理后的KBV200细胞,可使其p?Met蛋白的相对表达水平明显降低(P＜0.001).与二甲基亚砜(DMSO)组比较,AMG?102联合照射可明显增加Hep?2细胞的放射敏感性( SER＝1.28,P＜0.001);而AMG?102对KBV200细胞的放射敏感性影响很小(SER＝1.18,P＝0.002).与4 Gy照射组和5 μmol／L AMG?102处理组比较,4 Gy照射+5 μmol／L AMG?102处理组Hep?2细胞的凋亡率明显增加,cleaved caspase?3蛋白的表达水平明显升高.而各处理组 KBV200 细胞的凋亡率和 cleaved caspase?3蛋白的表达水平无明显变化.与DMSO处理组比较,4 Gy照射组、5 μmol／L AMG?102处理组和4 Gy照射+5 μmol／L AMG?102处理组Hep?2细胞中p?Met、p?Akt和p?Erk蛋白的表达水平下降,其中4 Gy照射+5 μmol／L AMG?102处理组中p?Met、p?Akt和p?Erk蛋白的表达水平比4 Gy照射组和5 μmol／L AMG?102处理组更低.但在KBV200细胞中,各处理组 |
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| AbstractList | 目的 探讨c?Met抑制剂AMG?102对喉鳞癌细胞的增殖抑制和放射增敏作用.方法 采用四甲基偶氮唑蓝(MTT)法检测AMG?102对喉鳞癌细胞株Hep?2和KBV200的增殖抑制作用.采用克隆形成实验分析AMG?102对Hep?2和KBV200细胞的放射增敏作用.采用流式细胞术检测Hep?2和KBV200细胞的凋亡情况.采用Western blot法检测细胞中c?Met／p?Met、凋亡蛋白cleaved caspase 3及下游信号通路蛋白Akt／p?Akt和Erk／p?Erk的表达.利用RNA干扰技术下调细胞中c?Met表达,转染c?Met过表达质粒使细胞中c?Met过表达,分析Hep?2和KBV200细胞对AMG?102的敏感性变化.结果 与KBV200细胞比较,Hep?2细胞对AMG?102更加敏感,半数抑制浓度(IC50)分别为14和9 μmol／L. Hep?2细胞中c?Met和p?Met蛋白的相对表达量分别为194.48±0.57和177.76±1.53,均明显高于KBV200细胞(分别为171.24±1.00和115.37±0.56,均P＜0.001). AMG?102作用于肝细胞生长因子(HGF)处理后的KBV200细胞,可使其p?Met蛋白的相对表达水平明显降低(P＜0.001).与二甲基亚砜(DMSO)组比较,AMG?102联合照射可明显增加Hep?2细胞的放射敏感性( SER＝1.28,P＜0.001);而AMG?102对KBV200细胞的放射敏感性影响很小(SER＝1.18,P＝0.002).与4 Gy照射组和5 μmol／L AMG?102处理组比较,4 Gy照射+5 μmol／L AMG?102处理组Hep?2细胞的凋亡率明显增加,cleaved caspase?3蛋白的表达水平明显升高.而各处理组 KBV200 细胞的凋亡率和 cleaved caspase?3蛋白的表达水平无明显变化.与DMSO处理组比较,4 Gy照射组、5 μmol／L AMG?102处理组和4 Gy照射+5 μmol／L AMG?102处理组Hep?2细胞中p?Met、p?Akt和p?Erk蛋白的表达水平下降,其中4 Gy照射+5 μmol／L AMG?102处理组中p?Met、p?Akt和p?Erk蛋白的表达水平比4 Gy照射组和5 μmol／L AMG?102处理组更低.但在KBV200细胞中,各处理组 |
| Abstract_FL | Objective To investigate the effect of c?Met inhibitor AMG?102 on proliferation and radiosensitivity in laryngeal squamous carcinoma cells. Methods The effects of AMG?102 on proliferation and radiosensitivity of laryngeal squamous carcinoma cell lines Hep?2 and KBV200 were detected by 3?(4,5?dimethy?2?thiazolyl)?2, 5?diphenyl?2H tetrazolium bromide ( MTT ) assay and colony formation assay, respectively. The apoptosis of Hep?2 and KBV200 cells was detected by flow cytometry.The expression levels of c?Met, phospho?Met (p?Met), cleaved caspase?3 and Akt／p?Akt, Erk／p?Erk were detected by Western blot. Specific small interfering RNA targeting c?Met or plasmid of c?Met were transfected into Hep?2 and KBV200 cells to investigate the cell sensitivity to AMG?102. Results Compared with KBV200 cells, Hep?2 cells were more sensitive to AMG?102 with IC50 of 14 and 9 μmol／L, respectively. The relative expression levels of c?Met and p?Met proteins in Hep?2 cells were 194.48±0.57 and 177.76±1.53, respectively, which were significantly higher than those in KBV200 cells ( 171.24 ± 1.00 and 115.37 ± 0.56, respectively, P＜0.001 for both). Exogenous hepatocyte growth factor (HGF) was added to increase the expression level of p?Met protein in KBV200 cells.The results showed that AMG?102 significantly reduced the expression of p?Met in KBV200 cells treated with HGF ( P＜0.001). Compared with the dimethyl sulfoxide ( DMSO) group, AMG?102 treatment combined with radiotherapy significantly increased the radiosensitivity of Hep?2 cells ( SER＝1.28, P＜0.001). However, AMG?102 had little effect on the radiosensitivity of KBV200 cells (SER＝1.18, P＝0.002). Compared with the 4 Gy radiotherapy alone group and the 5 μmol／L of AMG?102 alone treatment group, the apoptosis rate of Hep?2 cells in the combined treatment group was significantly increased. Meanwhile, the expression level of cleaved caspase?3 protein was also markedly increased. However, there were no significant changes in the apoptotic rate and cleaved caspase?3 expression in each treatment group of KBV200 cells. Compared with DMSO treatment group, the expression levels of p?Met, p?Akt and p?Erk were significantly decreased in the 4 Gy radiotherapy group, 5 μmol／L of AMG?102 treatment group and combined treatment group of Hep?2 cells. And the levels of p?Met, p?Akt and p?Erk in the combined treatment group were significantly lower than those in the 4 Gy radiotherapy alone group and 5 μmol／L of AMG?102 treatment alone group. By contrast, in KBV200 cells, the expression of p?Met, p?Akt and p?Erk in each group was not changed. The relative expression of p?Met in Hep?2 cells before and after radiotherapy at 30 min, 1 h, 4 h, 8 h, 24 h were 99.89±0.61, 138.62±1.00, 163.07±5.00, 87.80±1.85, 90.67±0.65 and 94.09±1.41, respectively. The level of p?Met was slightly increased after radiotherapy at 30 min and 1 h (P＜0.001 for all), whereas it was significantly decreased from 4 h to 24 h after radiotherapy (P＜0.05 for all). By contrast, the expression of p?Met in KBV200 cells did not change with time after radiotherapy (P＞0.05). The sensitivity of Hep?2 cells to AMG?102 was decreased after silencing of c?Met, while the sensitivity of KBV200 cells to AMG?102 was not significantly changed ( P＞0.05). Moreover, the radiosensitivity of Hep?2 cells in c?Met knockdown group had a slightly increasing trend ( SER＝1.07, P＝0.068). After the treatment with 10 μmol／L of AMG?102, the proliferation rate of c?Met ectopically expressed KBV200 cells was 60.05%± 3.23%, It was significantly lower than that of the blank control 90.08%±1.04% and siRNA negative control (90.12%±1.01%, P＜0.001). The results suggested that the overexpression of c?Met in KBV200 cells increased the radiosensitivity to AMG?102, whereas depletion of c?Met resulted in resistance to AMG?102 in Hep?2 cells. Furthermore, the radiosensitivity of KBV200 cells that overexpressed c?Met showed a decreased trend (SER＝0.7, P＝0.005). Conclusions c?Met inhibitor AMG?102 has a significant inhibitory effect on the proliferation of c?Met overexpressing laryngeal squamous carcinoma cells, leading to increased radiosensitivity. It suggests that molecular targeted therapy against c?Met receptor is more effective in c?Met overexpressed subtype of laryngeal squamous cell carcinoma. |
| Author | 马博敬 张建军 侯春立 吕欣 张翠红 范才 曹峰 陈坤 董凯峰 |
| AuthorAffiliation | 河北医科大学第四医院放疗科,石家庄,050011%邢台市柏乡县中心医院医务科055450%河北医科大学第一医院耳鼻喉科,石家庄,050031%解放军联勤保障部队第九八〇医院放疗科,石家庄,050082%解放军联勤保障部队第九八〇医院耳鼻喉头颈外科,石家庄,050082%解放军联勤保障部队第九八〇医院肿瘤科,石家庄,050082 |
| AuthorAffiliation_xml | – name: 河北医科大学第四医院放疗科,石家庄,050011%邢台市柏乡县中心医院医务科055450%河北医科大学第一医院耳鼻喉科,石家庄,050031%解放军联勤保障部队第九八〇医院放疗科,石家庄,050082%解放军联勤保障部队第九八〇医院耳鼻喉头颈外科,石家庄,050082%解放军联勤保障部队第九八〇医院肿瘤科,石家庄,050082 |
| Author_FL | Lyu Xin Zhang Cuihong Hou Chunli Zhang Jianjun Dong Kaifeng Fan Cai Chen Kun Ma Bojing Cao Feng |
| Author_FL_xml | – sequence: 1 fullname: Cao Feng – sequence: 2 fullname: Lyu Xin – sequence: 3 fullname: Dong Kaifeng – sequence: 4 fullname: Fan Cai – sequence: 5 fullname: Zhang Jianjun – sequence: 6 fullname: Chen Kun – sequence: 7 fullname: Ma Bojing – sequence: 8 fullname: Hou Chunli – sequence: 9 fullname: Zhang Cuihong |
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| DocumentTitle_FL | Effect of c?Met inhibitor AMG?102 on radiosensitivity in laryngeal squamous carcinoma cells |
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| Keywords | c?Met Cell proliferation 喉鳞状细胞癌 增殖 Radiosensitivity Laryngeal squamous cell carcinoma 凋亡 放射增敏 AMG?102 Apoptosis |
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| Publisher | 河北医科大学第四医院放疗科,石家庄,050011%邢台市柏乡县中心医院医务科055450%河北医科大学第一医院耳鼻喉科,石家庄,050031%解放军联勤保障部队第九八〇医院放疗科,石家庄,050082%解放军联勤保障部队第九八〇医院耳鼻喉头颈外科,石家庄,050082%解放军联勤保障部队第九八〇医院肿瘤科,石家庄,050082 |
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| Snippet | 目的 探讨c?Met抑制剂AMG?102对喉鳞癌细胞的增殖抑制和放射增敏作用.方法... |
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| Title | c-Met抑制剂AMG-102增强喉鳞癌细胞放射敏感性的机制 |
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