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、40 μg/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（20 μg/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\[(1.6±0.2)×104 vs （3.8±0.3)×104、(1.7±0.3)×104 vs (4.5±0.4)×104、 (2.5±0.1)×104 vs (4.0±0.4)×104、(1.5±0.1)×104 vs (2.5±0.3)×104个细胞, 均P<0.01\]，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 、40 μg/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 (\[1.6±0.2\]×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4 vs \[2.5±0.3\]×104, 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 G0/G1 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 (P<0.01); (4) CMSP significantly inhibited the proliferation and migration ability of TE-13 cells and induced cell differentiation; (5) the protein level of p-P38 was significantly increased while protein levels of p-ERK, p-SPKA/JNK were significantly decreased in TE-13 cells (P<0.01). Conclusion: CMSP suppressed TE-13 cell proliferation and induced the differentiation of TE-13 cells by up-regulating the level of p-P38 and down-regulating the levels of p-ERK, p-SPKA/JNK in MAPK signaling pathway.

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）