伊维菌素上调自噬水平抑制犬乳腺肿瘤细胞的增殖

Ivermectin Inhibits Canine Mammary Tumor Cell Growth by Upregulating Autophagy

为了探讨伊维菌素对犬乳腺肿瘤细胞的作用及可能机制,本研究采用CCK-8检测不同浓度伊维菌素对犬乳腺肿瘤细胞(CMT 7364,CIPp)增殖的影响;annexinⅤ-FITC/PI双染色检测伊维菌素对细胞凋亡的影响;转染pEGFP-LC3质粒,荧光显微镜下观察伊维菌素对自噬小体形成的影响;Western Blot检测伊维菌素处理后细胞中自噬标志分子LC3Ⅰ、LC3Ⅱ的表达水平。结果发现:伊维菌素可抑制犬乳腺肿瘤细胞的增殖,并呈现时间与剂量依赖性关系,CMT 7364和CIPp细胞24 h的IC50值分别为10.2μmol/L和10.76μmol/L;伊维菌素对细胞凋亡水平影响轻微,12μmol/L处理24 h和48 h后凋亡细胞比例均小于3%,各处理间无统计学差异(P>0.05);伊维菌素可诱导细胞浆内自噬小体的形成,促进LC3Ⅰ向LC3Ⅱ转化,诱导细胞自噬。表明伊维菌素通过诱导犬乳腺肿瘤细胞自噬发挥抗肿瘤作用。

To investigate the effect and the mechanism about how ivermectin inhibits the proliferation of canine mammary tumor cell lines. Cell viability was detected using CCK8 assay;cell apoptosis was detected by an annexin V-FITC/propidium iodide(PI) apoptosis detection kit using a FACS Calibur flow cytometer;cell autophagy was observed by fluorescence microscopy, the expressions of autophagy markers, LC3Ⅰand LC3Ⅱ, were detected by Western Blot. Ivermectin treatment impaired the cell viability of canine mammary tumor cell lines in a dose-and time-dependent manners, IC50 was 10.2 μmol/L in CMT 7364 and 10.76 μmol/L in CIPp with 24 h ivermectin treatment;ivermectin had few effect on cell spoptosis, the percentages of apoptosis cell were lower 3% with 12 μmol/L ivermectin for 24 h and 48 h, and no significant difference was observed between groups(P>0.05);autophagosomes and the expression of LC3Ⅱwere increased with ivermectin treatment. The growth inhibition of ivermectin in canine mammary tumor cell lines is attributed to its ability to induce autophagy.