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从乳腺癌患者中分离、鉴定癌细胞并进行增殖培养

Isolation, Characterization and Proliferation of Cancer Cells from Breast Cancer Patients.

作者信息

Widowati Wahyu, Heriady Yusuf, Laksmitawati Dian Ratih, Jasaputra Diana Krisanti, Wargasetia Teresa Liliana, Rizal Rizal, Perdana Fajar Sukma, Amalia Annisa, Arlisyah Annisa, Khoiriyah Zakiyatul, Faried Ahmad, Subangkit Mawar

机构信息

Medical Research Centre, Faculty of Medicine, Maranatha Christian University, Bandung, West Java, Indonesia.

Faculty of Medicine, Bandung Islamic University, Bandung, Indonesia.

出版信息

Acta Inform Med. 2018 Dec;26(4):240-244. doi: 10.5455/aim.2018.26.240-244.

Abstract

INTRODUCTION

The most cases of cancer death, which are in the first rank among cancers suffered by women is breast cancer. The breast cancer therapy for patients has been done, but still not optimal, so it is necessary to understand the mechanism of therapy in model cell of breast cancer.

AIM

This study aim to develop an isolation technique of breast cancer cell from patients as a cancer cell model.

MATERIAL AND METHODS

Breast cancer cell isolation is performed by enzymatic methods using the collagen I and hyaluronidase. Then, breast cancer cells were characterized using flow cytometry based on the CD44/CD24 expression where MDA-MB468 and MCF-7 breast cancer cell lines were used as positive controls. Estrogen receptor (ER), progesterone receptor (PR), p53, HER2, and Ki67 expression were assessed using an immunohistochemistry assay.

RESULT AND DISCUSSION

The morphology of cancer cells was fibroblast like cells on the day 7 after isolation. Isolated breast cancer cells expressed 95.33±0.47% of CD44/CD24 and human epidermal growth factor receptor 2 (HER2) low expressions. Isolation of breast cancer cells can use In-house enzymatic protocol. Isolated breast cancer showed the same expression as MDA-MB468 (CD44/CD24) and HER2 compared to MCF-7 cell lines (CD44/CD24).

CONCLUSION

These cells belonged to a basal type of breast carcinoma and expressed CD44/CD24, then isolated BCCs can be used as model cancer cells for further research.

摘要

引言

癌症死亡病例中,乳腺癌在女性所患癌症中位居首位。针对乳腺癌患者的治疗已经开展,但仍未达到最佳效果,因此有必要了解乳腺癌模型细胞中的治疗机制。

目的

本研究旨在开发一种从患者体内分离乳腺癌细胞作为癌细胞模型的技术。

材料与方法

采用胶原酶I和透明质酸酶通过酶解法进行乳腺癌细胞分离。然后,基于CD44/CD24表达,使用流式细胞术对乳腺癌细胞进行表征,其中MDA-MB468和MCF-7乳腺癌细胞系用作阳性对照。采用免疫组织化学分析法评估雌激素受体(ER)、孕激素受体(PR)、p53、HER2和Ki67的表达。

结果与讨论

分离后第7天,癌细胞形态呈成纤维细胞样。分离出的乳腺癌细胞表达95.33±0.47%的CD44/CD24,且人表皮生长因子受体2(HER2)低表达。乳腺癌细胞的分离可采用内部酶法方案。与MCF-7细胞系(CD44/CD24)相比,分离出的乳腺癌细胞与MDA-MB468(CD44/CD24)和HER2的表达相同。

结论

这些细胞属于基底型乳腺癌,表达CD44/CD24,因此分离出的乳腺癌细胞可作为模型癌细胞用于进一步研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cfd/6311122/f4e3cee13f90/AIM-26-240-g001.jpg

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