CAJ | 학술논문

目的:观察木鳖子醇提物(ethanol extract from cochinchina momordica seed, CMSEE)对黑素瘤B16细胞增殖的影响,初步探讨其作用机制.方法:MTT法和克隆集落形成实验检测CMSEE对小鼠黑素瘤B16细胞生长的影响,倒置相差显微镜和瑞氏-吉姆萨染色观察细胞形态学变化,流式细胞技术(FCM)检测B16细胞周期分布和凋亡率的变化,比色法检测CMSEE对B16细胞黑素生成和酪氨酸酶活性的影响,RT-PCR法检测CMSEE对B16细胞中C-myc、P38和Tyr基因表达的影响.结果:CMSEE(10～100 mg/L)明显抑制B16细胞的增殖(P<0.05, P<0.01),呈质量浓度和时间依赖性.10～40 mg/L CMSEE处理后,B16细胞呈现典型的细胞分化形态,细胞周期阻滞于G_0/G_1期,黑素产生和酪氨酸酶活性增加(P<0.01),C-myc基因表达下调,P38和Tyr基因表达上调;100 mg/LCMSEE处理后的B16细胞呈现凋亡形态变化[凋亡率达(37.2±3.29)%],黑素生成减少(P<0.05),酪氨酸酶活性降低(P<0.01).结论:CMSEE体外对B16细胞增殖有明显的抑制作用,其机制与其低剂量诱导分化和高剂量诱导凋亡有关.
목적:관찰목별자순제물(ethanol extract from cochinchina momordica seed, CMSEE)대흑소류B16세포증식적영향,초보탐토기작용궤제.방법:MTT법화극륭집락형성실험검측CMSEE대소서흑소류B16세포생장적영향,도치상차현미경화서씨-길모살염색관찰세포형태학변화,류식세포기술(FCM)검측B16세포주기분포화조망솔적변화,비색법검측CMSEE대B16세포흑소생성화락안산매활성적영향,RT-PCR법검측CMSEE대B16세포중C-myc、P38화Tyr기인표체적영향.결과:CMSEE(10～100 mg/L)명현억제B16세포적증식(P<0.05, P<0.01),정질량농도화시간의뢰성.10～40 mg/L CMSEE처리후,B16세포정현전형적세포분화형태,세포주기조체우G_0/G_1기,흑소산생화락안산매활성증가(P<0.01),C-myc기인표체하조,P38화Tyr기인표체상조;100 mg/LCMSEE처리후적B16세포정현조망형태변화[조망솔체(37.2±3.29)%],흑소생성감소(P<0.05),락안산매활성강저(P<0.01).결론:CMSEE체외대B16세포증식유명현적억제작용,기궤제여기저제량유도분화화고제량유도조망유관.
Objective: To investigate the effect of cochinchina momordica seed ethanol extract (CMSEE) on the proliferation of melanoma B16 cells and the underlying mechanism. Methods: MTT and clone formation assay were used to assess the effect of CMSEE on the growth of B16 cells. Morphological changes of B16 cells were observed under phase-contrast microscope and Giemsa staining. Cell cycle and apoptosis rate were examined by flow cytometry (FCM). The effect of CMSEE on melanin production and tyrosinase activity of B16 cells was assessed by colorimetry. The effect of CMSEE on the expression of C-myc, P38, and Tyr genes was examined by RT-PCR. Results: CMSEE (10-100 mg/L) inhibited the proliferation of B16 cell in a dose-and time-dependent manner. After treatment with 10-40 mg/L CMSEE, B16 cells showed typical differentiation morphology, and melanin production and tyrosinase activity were increased. B16 cells treated with 100 mg/L CMSEE showed apoptotic morphology, decreased melanin production and tyrosinase activity. B16 cell number in G_0/G_1 phase was significantly increased (P<0.01); C-myc mRNA expression was down-regulated, and P38, Try mRNA expression was up-regulated in B16 cells after treatment with 10-40 mg/L CMSEE. Conclusion: CMSEE can markedly inhibit the proliferation of melanoma B16 cells, which is related to induction of differentiation and promotion of apoptosis of B16 cells.