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Effect of CP metronomic chemotherapy on HUVEC cells and autophagy-related protein Beclin1 and LC3 |
GUO Lieping1, ZHOU Fan1, SHI Haotian1, CHEN Haimin1, LIN Chenhui1, CHEN Xiaoling1, HOU Jian1, 2 |
1. Department of Hematology and Oncology, Shanghai Zhabei District Central Hospital, Shanghai200070, China; 2. Department of Hematology, Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China |
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Abstract Objective To investigate the effect of CP metronomic chemotherapy on HUVEC cells and autophagy-related protein, Beclin1 and LC3, expression levels, and to explore the mechanism of CP program increasing sensitivity of HUVEC cells to chemotherapeutic drug. Methods Subjects were divided into the DMSO control group, and the phosphoramide mustard (PM) group, the prednisolone group, the phosphoramide mustard and prednisolone group (the CP group). HUVEC cells were treated with different drugs. CCK8 method was used to detect cell proliferation, flow cytometry to detect the cell apoptosis, Western blot and RT-PCR to detect autophagy-related protein levels. Results CCK8 method showed that compared with DMSO control group, in the PM, the prednisolone and the CP groups, HUVEC cell proliferation inhibition rates increased significantly with the extension of time (P<0.001). And at the same time, the different drug groups were significantly different in cell proliferation inhibition rate, the prednisone group was the weakest, the CP group(72 h,25.1%±0. 7%)was the strongest, and the PM group (72 h,20.4%±0.7%) was intermediate(P<0.01). Flow cytometry showed that after 48 hours, compared with the DMSO control group, the prednisolone, the PM and the CP groups made HUVEC cell apoptosis rate increased in turn (P<0.001). Western blot and reverse transcription PCR showed that after 48 hours, compared with the DMSO control group, Beclin1 and LC3 protein and mRNA expression decreased in turn in the prednisolone group, the PM group and the CP group(P<0.001). Conclusions HUVEC cells treated with CP metronomic chemotherapy can significantly reduce the level of HUVEC cell proliferation, promote HUVEC cell apoptosis, and significantly reduce autophagy-related protein expression levels of Beclin1 and LC3.
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Received: 25 November 2014
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Gozuacik D, Kimchi A. Autophagy as a cell death and tumor suppressor mechanism [J]. Oncogene, 2004, 23(16): 2891-2906.
|
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Holm E, Hildebrandt W, Kinsecherf R. Low postabsorptive net prtein degradation in male cancer patients: lack of sensitivity to regulatory amino acids [J]. Oncol Rep, 2007, 17(3): 695-700.
|
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付 俊, 尚海旭, 贾弘禔, 等. Beclin1与自噬及肿瘤的关系[J]. 生理科学进展, 2012, 43(2): 155-158.
|
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Satoh J, Motohashi N, Kino Y, et al. LC3, an autophagosome marker, is expressed on oligodendrocytes in Nasu-Hakola disease brains [J]. Orphanet J Rare Dis, 2014, 9: 68.
|
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黄豪博. VEGF反义RNA对骨髓瘤细胞U266生长及血管生成作用研究[D]. 福建医科大学, 2006.
|
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Sun Y, Liu J H, Sui Y X, et al. Beclin1 overexpression inhibitis proliferation, invasion and migration of CaSki cervical cancer cells [J]. Asian Pac J Cancer Prev, 2011, 12(5): 1269-1273.
|
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Guo X L, Li D, Sun K, et al. Inhibition of autophagy enhances anticancer effects of bevacizumab in hepatocarcinoma [J]. J Mol Med (Berl), 2013, 91(4): 473-483.
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