摘要
目的明确乳腺癌MCF-7细胞CD44+CD24-表达是否具有放化疗抵抗性,并研究其转移潜能相关性。方法无菌条件下通过FACS分选出CD44+CD24-MCF-7和non CD44+CD24-MCF-7两组细胞,进行同种条件下传代培养,采用5-氟尿嘧啶(5-FU)及医用X射线对两组细胞进行干预,对比检测两组细胞经干预后的生物学特性差异、体外侵袭能力及侵袭关联基因表达差异。MTT实验检测两组细胞经不同药物浓度的5-FU处理后细胞增殖率差异;流式细胞术检测经同种药物浓度的5-FU处理后两组细胞的周期及凋亡;彗星实验检测两组细胞经医用X射线照射后细胞DNA的损伤;蛋白质印迹法检测两组细胞侵袭相关基因PRDM14表达;基质胶侵袭性实验用于对比两组细胞体外侵袭能力差异。结果CD44+CD24-MCF-7细胞增殖抑制率为(93.86±2.95)%,与non CD44+CD24-MCF-7细胞(334.21±4.20)%比较明显下降,其IC50值约为后者4倍,u=0.38,P<0.01;细胞凋亡率CD44+CD24-MCF-7细胞为(8.77±0.66)%,较non CD44+CD24-MCF-7细胞(28.64±1.25)%显著降低,u=0.42,P=0.014;静止期(G0/G1期)CD44+CD24-MCF-7细胞为(76.19±1.623)%,较non CD44+CD24-MCF-7细胞(49.74±3.214)%明显增多,u=0.35,P=0.025;DNA损伤(4Gy Olive尾距值)CD44+CD24-MCF-7为13.130 0±0.542 8,较non CD44+CD24-MCF-7细胞33.250 0±0.932 7轻,P<0.000 1;体外侵袭能力CD44+CD24-MCF-7为75.28±6.932,较non CD44+CD24-MCF-7细胞24.23±3.408更强,P=0.007 6;PRDM14蛋白表达CD44+CD24-MCF-7为0.95±0.62,较non CD44+CD24-MCF-7细胞0.33±0.44上调,P<0.01。结论 CD44+CD24-MCF-7细胞相比non CD44+CD24-MCF-7细胞具有明显不同的细胞生物学特性,表明CD44+CD24-MCF-7表达细胞不仅具有明显的放化疗抵抗性,而且具有一定的转移潜能,在乳腺癌的化疗耐药或晚期转移中可能扮演一定的作用。
OBJECTIVE To clarify whether MCF-7 ceils with CD44＋ CD24- expression were resistant to chemo-ra- diotherapy and relevant to metastatic potential. METHODS Two groups of cells （CD44＋ CD24- cells and non CD44＋ CD24- cells） were sorted from the breast cancer cells lines MCF-7 by FACS under sterile conditions. Then this two groups of cells were cultured in the same condition. Biological characteristics of the cells were detected after the treatment of chemotherapy drugs and medical X-ray irradiations Its invasiveness capacity in vitro and the expression of invasiveness- related gene were further tested in the two group cells. MTT assay was introduced to detect the cell proliferation ratio af- ter different doses of 5-FU treatment; The cell cycle and cell apoptosis were detected by FCM; DNA breaks were detected by Comet assay after the irradiation of X-ray; Western blotting was used to detect the expression of invasion related genes PRDM14; Matrigel invasion experiments （Transwell assay） were used to compare the differences of invasion ability in the two group cells in vitro. RESULTS Compare to non CD44＋ CD24 MCF-7 cells, apoptosis rate （8.77 ± 0. 66）% vs （28.64± 1.25）% （p = 0. 014） was significantly reduced and proliferation rate, IC50： （334. 21 ±4. 20） % vs （ 93. 86 ±2.95） % （P〈0.01）was increased in CD44＋ CD24- MCF-7 cells; Quiescent cells increased significantly [Go/G1 ：（76.19± 1. 623）% vs （49.74±3. 214）% , （P= 0. 025）]; DNA breaks of CD44＋ CD24- MCF-7 cells were significantly decreased （4 Gy OTM value： 13. 130 0±0. 542 8 vs 33. 250 0i0. 932 7, P〈0. 000 1） % CD44＋ CD24- MCF-7 had a stronger inva- sive capability in vitro （75.28±6. 932 vs 24.23±3. 408,P=0. 007 6） and highly expressed PRDM14 protein （0.95± 0.62 vs O. 33vs0.44,P〈0.01）. CONCLUSIONS CD44＋ CD24- MCF-7 cells have quite different biological characteris- tics compared to CD44＋ CD24- MCF-7 cells, which reveals that MCF-7 cells not only have a signi{icant resistance
出处
《中华肿瘤防治杂志》
CAS
北大核心
2015年第13期1004-1008,1013,共6页
Chinese Journal of Cancer Prevention and Treatment
