| 孙岩岩,陈之显,李广恩,杨雨澎,张振龙,张晓亮.丹参酮通过VEGF/VEGFR信号通路抑制肝癌细胞迁移和侵袭能力的实验研究[J].,2020,(16):3033-3037 |
| 丹参酮通过VEGF/VEGFR信号通路抑制肝癌细胞迁移和侵袭能力的实验研究 |
| Experimental Study on Tanshinone Inhibiting Migration and Invasion Ability of Hepatoma Cells by VEGF/VEGFR Signaling Pathway |
| 投稿时间：2020-02-23 修订日期：2020-03-19 |
| DOI：10.13241/j.cnki.pmb.2020.16.007 |
| 中文关键词: 肝癌 丹参酮 细胞迁移 细胞侵袭 |
| 英文关键词: Hepatoma Tanshinone Cell migration Cell invasion |
| 基金项目:黑龙江省卫生计生委科研项目（2017-151）；哈尔滨医科大学创新科学研究项目（2017LCZX112） |
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| 中文摘要: |
| 摘要 目的：探究丹参酮通过VEGF/VEGFR信号通路抑制肝癌细胞迁移和侵袭能力的机制。方法：体外培养人肝癌细胞Hep3B、HepG2，并分为：实验组、阳性对照组和空白对照组，实验组使用丹参酮处理，阳性对照组使用阿霉素处理，空白对照为加入10 μL DMSO或生理盐水，使用CCK8检测肝癌细胞的增殖情况；分别加入浓度为31 μmol/L和2.5 μmol/L的丹参酮及阿霉素显微镜下观察细胞形态变化；使用流式细胞术检测肝癌细胞的凋亡情况和细胞周期情况；使用Western blot检测肝癌细胞VEGF及VEGFR蛋白表达情况。结果：MTT实验结果显示，随着丹参酮和阿霉素使用浓度的升高人肝癌细胞Hep3B、HepG2生长受到显著的抑制（P<0.05）；显微镜下观察发现丹参酮可以抑制肝癌肿瘤细胞增殖；流式细胞检测发现相比空白对照组，阳性对照组和实验组（人肝癌细胞Hep3B、HepG2）细胞凋亡率显著增加（P<0.05）；相比空白对照组，实验组和阳性对照组G0/G1期细胞百分比显著增加（P<0.05），S期和G2/M期细胞百分比显著降低（P<0.05）；蛋白质印记实验结果显示，相比空白对照组，实验组和阳性对照组VEGF及VEGFR蛋白表达显著降低（P<0.05），且实验组表达显著低于阳性对照组（P<0.05）。结论：参酮通可以通过抑制VEGF/VEGFR信号通路，将肝癌细胞分裂阻滞在G0/G1，达到抑制肝癌细胞的增殖及迁移和侵袭能力的效果。 |
| 英文摘要: |
| ABSTRACT Objective: To explore mechanism of tanshinone inhibiting migration and invasion ability of hepatoma cells by VEGF/VEGFR signaling pathway. Methods: Human hepatoma cells Hep3B and HepG2 were cultured in vitro. They were divided into experimental group, positive control group and blank control group. The experimental group was treated with tanshinone, positive control group was treated with doxorubicin, and blank control was given 10 μL DMSO or normal saline. CCK8 was applied to detect proliferation of hepatoma cells. 31 μmol/L and 2.5 μmol/L tanshinone and doxorubicin were applied. The changes of cell morphology were observed under microscope. The apoptosis and cycle of hepatoma cells were detected by flow cytometry. The expression of VEGF and VEGFR proteins was detected by Western blot. Results: The results of MTT assay showed that with increase of tanshinone and doxorubicin concentration, growth of human hepatoma cells Hep3B and HepG2 was significantly inhibited (P<0.05). It was found under microscope that tanshinone could inhibit proliferation of hepatocellular carcinoma cells. Flow cytometry test found that compared with the blank control group, apoptosis rates (human hepatoma cells Hep3B, HepG2) were significantly increased in positive control group and experimental group (P<0.05). Compared with blank control group, percentage of cells in G0/G1 phase was significantly increased in experimental group and positive control group (P<0.05), while percentage of cells in S phase and G2/M phase was significantly decreased (P<0.05). The results of Western blot showed that compared with blank control group, expression of VEGF and VEGFR proteins was significantly decreased in experimental group and positive control group (P<0.05). And the expression in experimental group was significantly lower than that in control group (P<0.05). Conclusion: Tanshinone can block hepatoma cell division in G0/G1 phase by inhibiting VEGF/VEGFR signaling pathway, thus inhibit proliferation, migration and invasion of hepatoma cells. |
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