多西他赛对黏液表皮样癌细胞系MC3中的肿瘤干细胞样细胞的影响  

Effect of docetaxel on cancer stem cell-like cells in the mucoepidermoid carcinoma cell line MC3

作　　者：张娄强[1] 王寅[1] 孙庚林[1] 

机构地区：[1]天津医科大学总医院口腔科,天津医学博士300052

出　　处：《医学研究生学报》2018年第2期152-157,共6页Journal of Medical Postgraduates

基　　金：天津市卫生局科技基金(2015KZ113)

摘　　要：目的多西他赛通常用于涎腺黏液表皮样癌术后化疗,但化疗并不能完全杀死残余的肿瘤细胞,肿瘤干细胞可能是肿瘤复发及耐药性的根源。文中基于肿瘤干细胞理论,探讨多西他赛对黏液表皮样癌细胞系MC3中的肿瘤干细胞样细胞的影响。方法取对数生长期的MC3细胞,分为阴性对照组(不加药物)、多西他赛组(加40 ng/m L多西他赛)。采用软琼脂克隆实验计算克隆形成率;MTT法检测细胞的生长;免疫细胞化学、流式细胞仪检测Oct4、CD44表达变化;裸鼠成瘤实验检测成瘤能力变化。结果多西他赛组MC3细胞克隆形成率[(33.47±1.30)%]较阴性对照组[(9.14±0.75)%]明显升高(P<0.05)。多西他赛组CD44+、Oct4+的表达[(99.50±0.30)%、(14.60±0.36)%]较阴性对照组[(14.47±0.15)%、(1.37±0.06)%]明显升高(P<0.05)。多西他赛组细胞的Oct4、CD44 mRNA的表达(2.228±0.005、0.651±0.015)明显高于阴性对照组(0.233±0.008、0.207±0.009),差异有统计学意义(P<0.05)。2组注射1×10~3个、1×10~4、1×10~5个细胞2个月后,多西他赛组分别未成瘤、1只裸鼠成瘤、3只裸鼠成瘤,而阴性对照组均未成瘤。结论多西他赛作用下存活的MC3细胞具有干细胞特性,多西他赛能够富集MC3细胞中的肿瘤干细胞,在肿瘤的复发中可能发挥重要作用。[ Abstract ] Objective Docetaxel is commonly used in chemotherapy for patients with mucoepidermoid carcinoma （MEC） in the salivary gland after operation, but it cannot kill all residual tumor cells and prevent recurrence. Cancer stem cells （CSCs） may be responsible for tumor recurrence and drug resistance. Based on the theory of CSCs, the authors investigated the effects of docetaxel on cancer stem cell-like cells in the MEC cell line MC3. Methods MC3 cells in the logarithmic growth phase were divided into a nega- tive control and a docetaxel （40 ng/mL） group. The colony formation rate of the MC3 cells was calculated with the soft agar cloning technique, the growth of the cells detected by MTT assay, the protein and mRNA expressions of CD44 and Oct4 determined by immu- nocytochemistry and flow cytometry, and tumorigenicity observed by nude mouse tumorigenicity assay. Results Compared with the negative control group, the MC3 cells treated with docetaxel exhibited significant increases in the colony formation rate （ [ 9.14±0.75 ] vs [ 33.47± 1.30] %, P〈0.05）, the protein expressions of CD44 （ 14.47±0.15 vs 99.50±0.30, P〈0.05） and Oct4 （ 1.37±0.06 vs 14.60±0.36, P〈0.05）, and the mRNA expressions of CD44 （0.207±0.009 vs 0.651±0.015, P〈0.05） and Oct4 （0.223±0.008 vs 2.228±0.005, P〈0.05）. At 2 months after injection of 1× 103, 1×104, and 1×105 MC3 cells, tumor formation was observed in 0, 1, and 3 of the nude mice, respectively, but not in the negative control group. Conclusion MC3 cells surviving docetaxel treatment have the properties of stem cells, and docetaxel can enrich cancer stem cells in the MC3 cells, which plays a key role in tumor recurrence.

关 键 词：肿瘤干细胞 黏液表皮样癌 OCT4 CD44 多西他赛 

分 类 号：R739.87[医药卫生—肿瘤]