Yudhani Ratih Dewi, Astuti Indwiani, Mustofa Mustofa, Indarto Dono, Muthmainah Muthmainah
Departement of Pharmacology, Faculty of Medicine, Sebelas Maret University, Surakarta, Indonesia. Email:
Departement of Pharmacology, Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia.
Asian Pac J Cancer Prev. 2019 Jun 1;20(6):1667-1673. doi: 10.31557/APJCP.2019.20.6.1667.
Background: Cervical cancer is one of the most prevalent gynecological cancers worldwide and contributes in high mortality of Indonesian women. The efficacy of chemotherapy as a standart therapy for cervical cancer decreases because it frequenly rises adverse effects. Recent studies have found that metformin has a potential anticancer effect mostly through reduction of cyclin expression and activation of Activated Adenosine Monophosphate Kinase (AMPK). This study aimed to investigate the effect of metfomin on expression of cyclin D1 and p53 and apoptosis in HeLa cancer cell line. Methods: HeLa cells were treated with various doses of metformin and doxorubicin as a positive control. Cytotoxic effect of metformin was determined using the MTT assay. Immunocytochemistry was used to assess cyclin D1 and p53 expression and apoptosis levels of treated HeLa cells were analyzed using flowcytometry. Data of cyclin D1 expression was statistically analyzed using the Kruskal-Wallis test followed by the Tamhane test, whilst ANOVA and Tukey post Hoc tests were used to analyze data of p53 and apoptosis level. The significant value was p< 0.05. Results: Metformin was able to inhibit proliferation of HeLa cells with IC50 60 mM. HeLa cells treated with 60 and 120 mM metformin had lower cyclin D1 expression than HeLa cells treated without metformin and reached a significant difference (p= 0.001). Moreover, 30 mM or higher doses of metformin increase significantly p53 expression (p< 0.001). Induction of apoptosis was observed in HeLa cells treated with all doses of metformin and reached statistically difference (p= 0.04 and p < 0.001). Conclusion: Metformin can modulate cyclin D1 and p53 expression in HeLa cancer cell line, leading to inhibition of cell proliferation and induction of apoptosis. Other cyclin family members, CDK inhibitors and AMPK signaling should be further investigated in order to know mechanism of metformin action.
宫颈癌是全球最常见的妇科癌症之一，也是导致印度尼西亚女性高死亡率的原因之一。化疗作为宫颈癌的标准治疗方法，其疗效因频繁出现不良反应而降低。最近的研究发现，二甲双胍具有潜在的抗癌作用，主要是通过降低细胞周期蛋白的表达和激活单磷酸腺苷活化蛋白激酶（AMPK）来实现的。本研究旨在探讨二甲双胍对HeLa癌细胞系中细胞周期蛋白D1和p53表达以及细胞凋亡的影响。方法：用不同剂量的二甲双胍处理HeLa细胞，并以阿霉素作为阳性对照。采用MTT法测定二甲双胍的细胞毒性作用。用免疫细胞化学法评估细胞周期蛋白D1和p53的表达，并用流式细胞术分析经处理的HeLa细胞的凋亡水平。细胞周期蛋白D1表达的数据采用Kruskal-Wallis检验，随后进行Tamhane检验进行统计学分析，而p53和凋亡水平的数据则采用方差分析和Tukey事后检验进行分析。显著性值为p<0.05。结果：二甲双胍能够抑制HeLa细胞的增殖，IC50为60 mM。用60 mM和120 mM二甲双胍处理的HeLa细胞，其细胞周期蛋白D1的表达低于未用二甲双胍处理的HeLa细胞，差异有统计学意义（p = 0.001）。此外，30 mM或更高剂量的二甲双胍能显著增加p53的表达（p<0.001）。在用所有剂量二甲双胍处理的HeLa细胞中均观察到凋亡诱导现象，差异有统计学意义（p = 0.04和p < 0.001）。结论：二甲双胍可调节HeLa癌细胞系中细胞周期蛋白D1和p53的表达，从而抑制细胞增殖并诱导细胞凋亡。为了解二甲双胍的作用机制，应进一步研究其他细胞周期蛋白家族成员、CDK抑制剂和AMPK信号通路。
