摘要
目的:探讨木鳖子单体化合物对羟基桂皮醛(p-hydroxylcinnamaldehyde,CMSP)对食管癌细胞株TE-13的分化作用及其机制。方法:用流式细胞术检测质量浓度为10、20、40Dμg/ml的CMSP处理TE-13细胞对该细胞凋亡和细胞周期分布的影响,用吉姆萨染色、电镜观察TE-13细胞形态学改变,Real-time PCR和ELISA方法检测TE-13细胞中肿瘤相关抗原CEA和SCC的表达,用克隆集落形成实验、Transwell迁移实验检测不同质量浓度CMSP对TE-13细胞的增殖、迁移能力的影响,Western blotting检测CMSP(20μg/ml)对TE-13细胞中MAPK通路中各蛋白水平的影响。结果:CMSP可抑制食管癌Kyse30、TE-13、Eca109和Kyse180细胞的增殖[(1.6±0.2)×10~4 vs(3.8±0.3)×10~4、(1.7±0.3)×10~4 vs(4.5±0.4)×10~4、(2.5±0.1)×10~4 vs(4.0±0.4)×10~4、(1.5±0.1)×10~4vs(2.5±0.3)×10~4个细胞,均P<0.01],而对人食管上皮细胞无明显作用(P>0.05)。CMSP处理TE-13细胞后:(1)随CMSP质量浓度(10、20、40μg/ml)和处理时间的增加(24、48、72 h),G_0/G_1期细胞数量均增加、S期细胞数量减少(P<0.01或P<0.05),但细胞凋亡率无明显变化(P>0.05);(2)细胞出现典型分枝状突起,且随CMSP质量浓度增加更显著(P<0.05);(3)CEA和SCC水平显著降低(P<0.01);(4)抑制TE-13细胞的增殖和迁移能力,诱导细胞分化;(5)p-P38蛋白水平明显升高(P<0.01),而p-ERK、P-SPKA/JNK蛋白水平明显降低(P<0.01)。结论:CMSP抑制食管癌TE-13细胞的增殖、诱导其分化,其作用机制可能与上调MAPK信号通路中P-P38和下调P-ERK、P-SPKA/JNK有关。
Objective：To investigate the effects of p-hydroxylcinnamaldehyde （CMSP）, a novel compound extracted from cochinchina momordica seed, on differentiation of esophageal carcinoma TE-13 cells and its possible mechanism. Methods： FCM was used to evaluate the effect of CMSP （at concentrations of 10, 20, 40 μg/ml） on the apoptosis and cell cycle distribution of TE-13 cells. Wright-Giemsa staining and electron microscope were used to observe the morphological changes of TE-13 cells. RT-qPCR and ELISA were used to evaluate the expressions of tumor associated antigens CEA（carcino-embryonic antigen）and SCC（squamous cell carcinoma antigen）in TE-13 cells after the treatment with CMSP. Influence of different concentrations of CMSP on proliferation and migration of TE-13 cells were determined by colony forming assay and transwell assay. Western blotting was used to evaluate the expression of the proteins in MAPK pathway of TE-13 cells that treated by CMSP （20 μg/ml）. Results： CMSP significantly inhibited the growth of esophagus cancer cell lines Kyse30, Te-13, Eca109 and Kyse180 （／[1.6±0.2／]×104 vs ／[3.8±0.3／]×104, ／[1.7±0.3／]×104 vs ／[4.5±0.4／]×104, ／[2.5±0.1／]×104 vs ／[4.0±04／]×104, ／[1.5±0.1／]×104 vs ／[2.5±0.3／]×104, all P〈0.01）, but had no significant effect on normal esophageal epithelial cells （P〉0.05）. After the treatment with CMSP, （1） the number of TE-13 cells at G0/G1 phase increased in a dose （10, 20, 40 μg/ml） and time （24, 48, 72 h）dependent manner （P〈0.01,P〈0.05）, while the number of cells at S phase decreased （P〈0.01, P〈0.05）; however, the apoptosis rate showed no obvious change （P〉0.05）; （2） TE-13 cells showed typical dendrite-like cellular protrusions, and the percentage of such elongated cells was significantly and progressively increased with the increase in CMSP concentration （P〈0.05）; （3） the expressions of CEA and SCC in TE-13 cells significantly decreased
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2017年第1期30-37,共8页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金青年基金资助项目(No.81502032)
河北省医学科学研究重点课题计划项目(No.20120120)~~
