香加皮杠柳苷通过溶酶体途径诱导胃癌MGC-803细胞凋亡

李磊; 赵连梅; 崔雯萱; 戴素丽; 彭克楠; 孙士萍; 张超; 单保恩

doi:10.3971/j.issn.1000-8578.2016.05.004

香加皮杠柳苷通过溶酶体途径诱导胃癌MGC-803细胞凋亡

Periplocin Extracted from Cortex Periplocae(CPP) Induced Apoptosis of Gastric Cancer Cells MGC-803 Through Lysosomal Pathway

摘要

摘要:
目的探讨香加皮杠柳苷（periplocin extracted from cortex periplocae, CPP）是否通过溶酶体途径诱导胃癌细胞凋亡。
方法 200、100、50 ng/ml香加皮杠柳苷作用于MGC-803细胞24和48 h后，MTS法检测细胞的增殖情况，用TUNEL法检测细胞凋亡，Western blot检测不同浓度香加皮杠柳苷作用于MGC-803细胞24 h后Caspase-3和Caspase-9的表达情况，吖啶橙染色检测溶酶体释放情况，免疫荧光法检测组织蛋白酶B（Cathepsin B）的表达变化情况。
结果与空白对照组相比，50、100、200 ng/ml香加皮杠柳苷作用于MGC-803细胞24、48 h后，均具有增殖抑制作用，24 h增殖率分别为 (66.43±1.67)%、(55.39±3.63)%、(37.06±0.57)% vs. (100.14±8.49)%，P均＜0.05，48 h增殖率分别为(38.16±3.26)%、(32.00±1.83)%、(24.75±1.57)% vs. (100.33±8.98)%，P均＜0.05。与空白对照组相比，50、100、200 ng/ml香加皮杠柳苷作用于MGC-803细胞24 h后，细胞凋亡率显著增加，凋亡率为(28.56±6.96)%、(33.70±4.60)%、(60.42±4.48)% vs. (2.23±1.03)%，P均＜0.05，Caspase-3的活性裂解片段和Caspase-9表达量升高，溶酶体膜的完整性被破坏，Cathepsin B由溶酶体释放到细胞质中。
结论香加皮杠柳苷可通过溶酶体途径诱导胃癌细胞凋亡。

Abstract:
Objective To investigate whether periplocin extracted from cortex periplocae (CPP) induces the apoptosis of gastric cancer cells through the lysosomal pathway.
Methods MGC-803 cells were treated with CPP at different concentrations(50, 100, 200ng/ml) for 24 and 48h. MTS assays were performed to measure the cell proliferation. The apoptosis of cells were analyzed using TUNEL. Western blot was used to examine the change of Capase-3 and Caspase-9 expression in MGC-803 cells treated with CPP at different concentrations for 24h. Acridine orange staining and immunofluorescence were introduced to observe the situation of lysosomes release and the change of Cathepsin B expression.
Results Compared with the blank control group (0ng/ml), CPP(50, 100, 200ng/ml) inhibited MGC-803 cells proliferation and induced cell apoptosis after 24 and 48h: the proliferation rates after 24h were (66.43±1.67)%, (55.39±3.63)%, (37.06±0.57)% vs. (100.14±8.49)% (P＜0.05), and after 48h were (38.16±3.26)%, (32.00±1.83)%, (24.75±1.57)% vs. (100.33±8.98)% (P＜0.05); the apoptosis rates after 24h were (28.56±6.96)%, (33.70±4.60)%, (60.42±4.48)% vs. (2.23±1.0)% (P＜0.05). The expression of cleaved Caspase-3 and Caspase-9 were elevated. The integrity of the lysosomal membrane was damaged and Cathepsin B was released to cytoplasm from lysosome.
Conclusion CPP could induce the apoptosis of gastric cancer cells through the lysosomal pathway.

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