CAJ | 학술논문

目的探讨甲基硒酸(MSA)对人三阴性乳腺癌(TNBC)的化疗增敏作用及其分子机制.方法分别采用2.5、3.5、4.0 μmol/L的MSA联合不同浓度的紫杉醇、阿霉素与MDA-MB-231细胞株共培养,分别应用细胞计数试剂盒(CCK-8)检测化疗药物单药和联合MSA用药时细胞增殖抑制率,并通过合用指数的计算,探讨MSA对化疗药物疗效的影响；应用膜联蛋白V-异硫氰酸荧光素(Annexin V-FITC)/碘化丙锭(PI)双染法检测细胞凋亡变化；利用Western blot检测蛋白去乙酰化-6(HDAC-6)的表达.结果不同浓度化疗药物联合MSA后细胞增殖率较单用化疗药物组均下降,呈明显的量效关系,提示MSA与化疗药物具有协同作用；MSA、紫杉醇及阿霉素对TNBC细胞均有诱导凋亡作用,10nmol/L紫杉醇联合3.5 μmol/L的MSA后细胞凋亡率由42.7％上升至60.4％(P＜0.01),0.5μmol/L阿霉素联合3.5 μmol/L的MSA后细胞凋亡率由31.6％上升至50.8％(P＜0.05)；单用化疗药物不影响HDAC-6的表达,MSA单药及联合化疗药物处理后细胞HDAC-6的表达受到明显抑制.结论 MSA通过增强抗肿瘤药物诱导细胞凋亡作用提高了化疗药物的疗效,调节HDAC-6活性是其化疗增敏的机制之一.
목적 탐토갑기서산(MSA)대인삼음성유선암(TNBC)적화료증민작용급기분자궤제.방법 분별채용2.5、3.5、4.0 μmol/L적MSA연합불동농도적자삼순、아매소여MDA-MB-231세포주공배양,분별응용세포계수시제합(CCK-8)검측화료약물단약화연합MSA용약시세포증식억제솔,병통과합용지수적계산,탐토MSA대화료약물료효적영향；응용막련단백V-이류청산형광소(Annexin V-FITC)/전화병정(PI)쌍염법검측세포조망변화；이용Western blot검측단백거을선화-6(HDAC-6)적표체.결과 불동농도화료약물연합MSA후세포증식솔교단용화료약물조균하강,정명현적량효관계,제시MSA여화료약물구유협동작용；MSA、자삼순급아매소대TNBC세포균유유도조망작용,10nmol/L자삼순연합3.5 μmol/L적MSA후세포조망솔유42.7％상승지60.4％(P＜0.01),0.5μmol/L아매소연합3.5 μmol/L적MSA후세포조망솔유31.6％상승지50.8％(P＜0.05)；단용화료약물불영향HDAC-6적표체,MSA단약급연합화료약물처리후세포HDAC-6적표체수도명현억제.결론 MSA통과증강항종류약물유도세포조망작용제고료화료약물적료효,조절HDAC-6활성시기화료증민적궤제지일.
Objective To investigate the effects of methylseleninic acid on chemosensitization of triple-negative breast cancer (TNBC) cells and the molecular mechanism.Methods Methylseleninic acid (MSA) (2.5,3.5,4.0 μmol/L) in combination with paclitaxel,doxorubicin or malondialdehyde (MDA)-treated MB-231 cells were cultured separately.Cell counting kit-8 (CCK-8) assay was applied to detect the inhibition of cell proliferation rate by chemotherapy drugs use alone or in combination with MSA,and the combined index was calculated to explore the impact of the function of MSA on the efficacy of chemotherapy drugs.Annexin V-FITC/propidium iodide (PI) double staining was applied to detect apoptosis,flow cytometry to examine cell cycle,and Western blotting to analyze thes histone deacetylase-6 (HDAC-6) expression.Results The cell proliferation rate of chemotherapy drugs in combination with MSA was decreased as compared with chemotherapy drugs used alone,suggesting the synergetic effects of MSA and chemotherapy drugs.The combined use of paclitaxel with MSA could significantly increase the number of cells in G2/M phase (P ＜ 0.05),that of doxorubicin with MSA could significandy increase the number of cells in S-phase cells (P ＜ 0.05),and the prompt MSA enhanced the effect of anticancer drugsinduced cell cycle arrest.MSA,paclitaxel and doxorubicin could induce apoptosis of TNBC cells.The apoptosis rate in MSA + chemotherapy groups was significantly increased as compared with monotherapy group and the control group.Chemotherapy drugs alone did not affect the expression of HDAC-6,and MSA used alone or incombination with chemotherapy drugs could significantly inhibit the HDAC-6 expression.Conclusion MSA enhanced anticancer drugs-induced cell cycle arrest and apoptosis so as to improve the efficacy of chemotherapy drugs probably by regulating the HDAC-6 expression.