Jing Xiaoping, Xu Yichun, Cheng Weiwei, Guo Sheng, Zou Ya, He Li
Department of Traditional Chinese Medicine, Shanghai Children's Hospital, Shanghai Jiao Tong University, 1400 West Beijing Road, Lane 24, Shanghai, Zip Code 200040, China.
Shanghai Biochip Co., Ltd, National Engineering Center for Biochip at Shanghai, Shanghai, 201203, China.
Cancer Chemother Pharmacol. 2016 Jun;77(6):1171-81. doi: 10.1007/s00280-016-3034-6. Epub 2016 Apr 21.
To investigate the occurrence of apoptosis and autophagy on human gastric cancer cells after treatment by Tanshinone I, as well as the relationship between them.
BGC823 and SGC7901 cells were treated with Tanshinone I; the cell proliferation was measured using CCK-8 and clone formation assay; and the expression of apoptosis- and autophagy-associated proteins was detected by Western blot. Autophagic vacuoles in cells were observed with LC3 dyeing using confocal fluorescent microscopy, and apoptotic cells were detected via flow cytometry. Bcl-2 was overexpressed in gastric cells treated with Tanshinone I or not, and autophagy relative protein was investigated; the interaction between Beclin-1 and Bcl-2 was detected by immunoprecipitation. Cell apoptosis was detected when autophagy was inhibited by ATG7-siRNA. Tumor growth was assessed by subcutaneous inoculation of cells into BALB/c nude mice.
Tanshinone I inhibited the proliferation of BGC823 and SGC7901 cells, and induced cell apoptosis by inhibiting anti-apoptosis protein Bcl-2. Tanshinone I also increased the conversion of LC3I to LC3II and triggered autophagosome formation, without changing the expression of Beclin-1. However, the Beclin-1 VPS34 complexes were increased after Tanshinone I treatment via inhibiting Bcl-2 expression. Moreover, disturbing autophagy by knockdown of ATG7 expression contributed to Tanshinone I-induced apoptosis. In vivo assay showed that combination with autophagy inhibitor chloroquine in nude mice bearing BGC823 xenograft significantly augmented the antitumor effect of Tanshinone I.
Tanshinone I induced apoptosis and pro-survival autophagy via inhibiting Bcl-2 expression on gastric cancer, and the combination of chloroquine and Tanshinone I could inhibit tumor growth more efficiently than monotherapy, which might be considered as an effective strategy for the treatment for gastric cancer.
研究丹参酮I处理后人胃癌细胞凋亡和自噬的发生情况及其二者之间的关系。
用丹参酮I处理BGC823和SGC7901细胞；采用CCK-8法和克隆形成实验检测细胞增殖；通过蛋白质免疫印迹法检测凋亡和自噬相关蛋白的表达。利用共聚焦荧光显微镜对细胞进行LC3染色观察自噬泡，通过流式细胞术检测凋亡细胞。在丹参酮I处理或未处理的胃癌细胞中过表达Bcl-2，并研究自噬相关蛋白；通过免疫沉淀检测Beclin-1与Bcl-2之间的相互作用。当自噬被ATG7-siRNA抑制时检测细胞凋亡。将细胞皮下接种到BALB/c裸鼠体内评估肿瘤生长情况。
丹参酮I抑制BGC823和SGC7901细胞的增殖，并通过抑制抗凋亡蛋白Bcl-2诱导细胞凋亡。丹参酮I还增加了LC3I向LC3II的转化并触发自噬体形成，而不改变Beclin-1的表达。然而，丹参酮I处理后通过抑制Bcl-2表达增加了Beclin-1 VPS34复合物。此外，通过敲低ATG7表达干扰自噬促进了丹参酮I诱导的凋亡。体内实验表明，在携带BGC823异种移植瘤的裸鼠中联合使用自噬抑制剂氯喹可显著增强丹参酮I的抗肿瘤作用。
丹参酮I通过抑制胃癌细胞中Bcl-2的表达诱导凋亡和促生存自噬，氯喹与丹参酮I联合使用比单一疗法更能有效抑制肿瘤生长，这可能是一种有效的胃癌治疗策略。
