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CD146 阳性牙髓来源干细胞和 CD146 阴性成纤维细胞中耐药基因的差异表达。

Differential expression of drug resistance genes in CD146 positive dental pulp derived stem cells and CD146 negative fibroblasts.

机构信息

Oral and Dental Disease Research Center, Department of Operative Dentistry, School of Dentistry, Shiraz Universityof Medical Sciences, Shiraz, Iran.

Department of Cardiovascular Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

出版信息

Clin Exp Dent Res. 2020 Aug;6(4):448-456. doi: 10.1002/cre2.297. Epub 2020 May 7.

Abstract

INTRODUCTION

The stem cell portion of the dental pulp derived cultures (DPSCs) showed a higher resistance to cytotoxic effect of restorative dental materials compared to pulpal fibroblasts (DPFs). Here, we aimed to compare the expression of some drug resistant genes between these cells.

METHODS AND MATERIALS

To separate DPSCs from DPFs, we used magnetic cell sorting technique based on CD146 expression. To assess the stem cell properties, the positive and negative portions underwent colony forming assays and were induced to be differentiated into the adipocytes, osteoblasts, hepatocytes, and neural cells. Cell surface antigen panels were checked using immune fluorescence and flow-cytometry techniques. The mRNA expression of 14 ABC transporters including ABCA2, ABCB1, ABCB11, ABCC1, ABCC2, ABCC3, ABCC4, ABCC5-2, ABCC5-4,ABCC5-13, ABCC6, ABCC10, ABCC11, and ABCG2 genes was assessed, using quantitative RT-PCR technique.

RESULTS

Only the CD146 positive portion could be differentiated into the desired fates, and they formed higher colonies (16.7 ± 3.32 vs. 1.7 ± 1.67, p < .001). The cell surface antigen panels were the same, except for CD146 and STRO-1 markers which were expressed only in the positive portion. Among the ABC transporter genes studied, the positive portion showed a higher expression (approximately two-fold) of ABCA2, ABCC5-13, and ABCC5-2 genes.

CONCLUSION

Dental pulp stem cells which can be separated from dental pulp fibroblasts based on CD146 expression, express higher levels of some drug resistance genes which probably accounts for their features of more resistance to cytotoxic effects of some dental materials. This needs to be more validated in future.

摘要

简介

与牙髓成纤维细胞(DPFs)相比,牙髓衍生培养物(DPSCs)中的干细胞部分对修复性牙科材料的细胞毒性作用具有更高的抵抗力。在这里,我们旨在比较这两种细胞之间一些耐药基因的表达。

方法和材料

我们使用基于 CD146 表达的磁细胞分选技术从 DPFs 中分离 DPSCs。为了评估干细胞特性,阳性和阴性部分进行了集落形成测定,并被诱导分化为脂肪细胞、成骨细胞、肝细胞和神经细胞。使用免疫荧光和流式细胞术技术检查细胞表面抗原谱。使用定量 RT-PCR 技术评估 14 种 ABC 转运蛋白(ABCA2、ABCB1、ABCB11、ABCC1、ABCC2、ABCC3、ABCC4、ABCC5-2、ABCC5-4、ABCC5-13、ABCC6、ABCC10、ABCC11 和 ABCG2 基因)的 mRNA 表达。

结果

只有 CD146 阳性部分才能分化为所需的命运,并且它们形成更高的集落(16.7±3.32 对 1.7±1.67,p<0.001)。细胞表面抗原谱相同,除了仅在阳性部分表达的 CD146 和 STRO-1 标记物。在所研究的 ABC 转运蛋白基因中,阳性部分表现出更高的表达(约两倍)的 ABCA2、ABCC5-13 和 ABCC5-2 基因。

结论

可以根据 CD146 表达从牙髓成纤维细胞中分离的牙髓干细胞表达更高水平的一些耐药基因,这可能解释了它们对某些牙科材料细胞毒性作用更具抵抗力的特征。这需要在未来进行更多验证。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d4d/7453779/f24b241be7d6/CRE2-6-448-g001.jpg

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