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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 differentiation of esophageal carcinoma cell line TE-13 induced by p-hydroxylcinnamaldehyde (CMSP) and its mechanism. Methods: The effect of CMSP on the apoptosis and cell cycle distribution of TE-13 cells treated with CMSP at concentrations of 10, 20 and 40 μg / ml was detected by flow cytometry. The morphology of TE-13 cells was observed by Giemsa staining and electron microscopy Real-time PCR and ELISA were used to detect the expression of tumor-associated antigen CEA and SCC in TE-13 cells. The effects of different concentrations of CMSP on the proliferation and migration of TE-13 cells were detected by colony formation assay and Transwell migration assay The effect of CMSP (20μg / ml) on the protein levels of MAPK pathway in TE-13 cells was detected by Western blotting. Results: CMSP could inhibit the proliferation of esophageal 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) × 10-4vs (2.5 ± 0.3) × 10-4 cells , Both P <0.01], but had no significant effect on human esophageal epithelial cells (P> 0.05). After CMSP treatment of TE-13 cells: (1) With the increase of CMSP concentration (10, 20 and 40μg / ml) and treatment time (24,48 and 72 h), the number of G_0 / (P <0.01 or P <0.05), but the apoptosis rate did not change significantly (P> 0.05). (2) The cells appeared typical dendrites and were more significant with the concentration of CMSP (P <0.05) ; (3) The levels of CEA and SCC were significantly decreased (P <0.01); (4) The proliferation and migration of TE-13 cells were inhibited and the differentiation was induced; (5) The level of p- , While the levels of p-ERK and P-SPKA / JNK were significantly decreased (P <0.01). CONCLUSION: CMSP can inhibit the proliferation and induce the differentiation of esophageal carcinoma TE-13 cells. The mechanism may be related to up-regulation of P-P38 and down-regulation of P-ERK and P-SPKA / JNK in MAPK signaling pathway.