灵菌红素对耐阿霉素人乳腺癌细胞MCF-7/ADR的作用研究

Study on the effect of Prodigiosin on adriamycin-resistant human breast cancer cells MCF-7/ADR

目的探讨灵菌红素对耐阿霉素人乳腺癌细胞MCF-7/ADR的作用。方法采用平板发酵粘质沙雷氏菌WA12-1-18生产灵菌红素,CCK-8测定灵菌红素的体外活性及MCF-7/ADR的耐药指数。构建裸鼠皮下移植瘤模型,灵菌红素高、低剂量组分别腹腔注射5.0、2.5 mg/kg灵菌红素,生理盐水对照位腹腔注射等体积生理盐水,每4天1次,连续用药24 d,观察移植瘤的体积、质量及裸鼠体质量,HE染色观察移植瘤组织及裸鼠主要脏器的病理情况,免疫组织化学检测移植瘤Ki-67的表达。结果获得的灵菌红素质量分数为95.18%,对MCF-7和MCF-7/ADR的IC50分别为0.484μg/mL和0.264μg/mL。MCF-7/ADR耐药指数13,符合细胞耐药株的要求。灵菌红素处理组裸鼠移植瘤增长速度较生理盐水组慢,肿瘤细胞排列稀疏,核小且着色较浅,Ki-67染色后棕色明显减少。灵菌红素2.5 mg/kg组和灵菌红素5 mg/kg组,在荷MCF-7/ADR裸鼠中抑瘤率分别为37.23%和53.72%。另外,各组裸鼠体质量差异无统计学意义,主要脏器无明显病理变化。结论灵菌红素可抑制裸鼠体内耐阿霉素人乳腺癌细胞MCF-7/ADR的生长,对心、肝、脾、肺和肾等主要脏器无明显毒副作用,为灵菌红素在耐阿霉素乳腺癌的临床应用提供了实验依据。

Objective To investigate the effect of Prodigiosin on adriamycin-resistant human breast cancer cell line MCF-7/ADR.Methods Prodigiosin was produced by plate fermentation of Serratia marcescens WA12-1-18.In vitro activity of Prodigiosin and the drug resistance index of MCF-7/ADR were determined by CCK-8.A subcutaneous xenograft tumor model was established in nude mice,and Prodigiosin with 5.0,2.5 mg/kg or equal volume saline were administered through intraperitoneal injection in high and low dose groupsɑnd normal saline control group once every 4 days for 24 days.The tumor volume,weight,and nude mouse body mass were observed.HE staining was used to observe the pathological conditions of transplanted tumor tissue and major organs in nude mice.Immunohistochemistry was used to detect the expression of Ki-67 in transplanted tumors.Results The purity of Prodigiosin was 95.18%,and the IC50 of Prodigiosin to MCF-7 and MCF-7/ADR were 0.484μg/mL and 0.264μg/mL,respectively.The drug resistance index of MCF-7/ADR was 13,which was in line with the requirement of drug-resistant cell strains.The growth rate of transplanted tumors in nude mice treated with Prodigiosin was slower than that in the saline group,with sparse arrangement of tumor cells,small nuclei,and lighter staining.After Ki-67 staining,the brown color significantly decreased.The tumor inhibition rates of the 2.5 mg/kg and 5 mg/kg groups of Prodigiosin in MCF-7/ADR bearing nude mice were 37.23%and 53.72%,respectively.In addition,there was no significant difference in body weight among the groups of nude mice,and there were no significant pathological changes in the main organs.Conclusion Prodigiosin can inhibit the growth of adriamycin-resistant human breast cancer cell MCF-7/ADR in nude mice without obvious toxic and side effects,which provides an experimental basis for the clinical application of Prodigiosin in adriamycin-resistant human breast cancer.