Expanding the prostate cancer cell line repertoire with ACRJ-PC28, an AR-negative neuroendocrine cell line derived from an African-Caribbean patient.
Expanding the prostate cancer cell line repertoire with ACRJ-PC28, an AR-negative neuroendocrine cell line derived from an African-Caribbean patient.
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DOI：
10.1158/2767-9764.crc-22-0245
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发表时间：
2022-11
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来自不同背景的前列腺癌细胞系对于解决黑人男性前列腺癌发病率和死亡率的差异非常重要。ACRJ-PC 28是从经直肠穿刺活检中开发的，并通过CDKN 2A基因座的失活和人端粒酶的同时表达来建立。表征分析包括生长曲线分析、免疫印迹、IHC、三维培养、免疫荧光成像、共聚焦显微镜、流式细胞术、全基因组测序（WGS）和RNA测序（RNA-seq）。ACRJ-PC 28在体外传代超过40次，时间超过10个月，倍增时间为45小时。短串联重复序列分析证实了该细胞系的新奇和人类起源。RNA-seq证实了前列腺特异性基因α-甲酰基-CoA消旋酶（AMACR）和NKX3.1的表达，神经内分泌特异性标志物突触素和烯醇化酶2（ENO 2）和IHC证实了AMACR的存在。免疫印迹表明该细胞系为基底腔型，表达p53和pRB，雄激素受体（AR）阴性。WGS证实了不存在外显子突变和存在内含子变体，这些变体似乎不影响AR、p53和pRB的功能。RNA-seq数据揭示了许多TP 53和RB 1 mRNA剪接变体以及AR mRNA表达的缺乏。这与响应于DNA损伤的p53功能和响应于接触抑制的pRB功能的保留一致。软琼脂贴壁独立分析表明细胞发生转化，通过主成分分析证实，其中ACRJ-PC 28细胞与其他前列腺癌肿瘤组织一起聚集，但不同。所描述的新方法应该推进前列腺细胞系的开发，解决黑人男性前列腺癌的差异。细胞系开发仍然吸引不到10%的成功率。超过98%的前列腺癌细胞系来自白色男性。这可能导致与患有前列腺癌的白色男性相比，患有前列腺癌的黑人男性对治疗的反应较差，从而增加了白色男性的总体生存率。本文描述的开发ACRJ-PC 28的方法应该促进黑色前列腺癌细胞系的存在，从而解决前列腺癌差异。
Prostate cancer cell lines from diverse backgrounds are important to addressing disparities in prostate cancer incidence and mortality rates among Black men. ACRJ-PC28 was developed from a transrectal needle biopsy and established via inactivation of the CDKN2A locus and simultaneous expression of human telomerase. Characterization assays included growth curve analysis, immunoblots, IHC, three-dimensional cultures, immunofluorescence imaging, confocal microscopy, flow cytometry, whole-genome sequencing (WGS), and RNA sequencing (RNA-seq). ACRJ-PC28 has been passaged more than 40 times in vitro over 10 months with a doubling time of 45 hours. Short tandem repeat profiling confirmed the novelty and human origin of the cell line. RNA-seq confirmed the expression of prostate specific genes alpha-methylacyl-CoA racemase (AMACR) and NKX3.1 and neuroendocrine specific markers synaptophysin and enolase 2 (ENO2) and IHC confirmed the presence of AMACR. Immunoblots indicated the cell line is of basal-luminal type; expresses p53 and pRB and is androgen receptor (AR) negative. WGS confirmed the absence of exonic mutations and the presence of intronic variants that appear to not affect function of AR, p53, and pRB. RNA-seq data revealed numerous TP53 and RB1 mRNA splice variants and the lack of AR mRNA expression. This is consistent with retention of p53 function in response to DNA damage and pRB function in response to contact inhibition. Soft agar anchorage-independent analysis indicated that the cells are transformed, confirmed by principal component analysis where ACRJ-PC28 cells cluster alongside other prostate cancer tumor tissues, yet was distinct. The novel methodology described should advance prostate cell line development, addressing the disparity in prostate cancer among Black men. Cell line development continues to attract less than 10% success rate. More than 98% of prostate cancer cell lines are from White men. This may contribute to the poorer response by Black men with prostate cancer to therapy compared with White men with prostate cancer, increasing overall survivorship among White men. The methodology described here to develop ACRJ-PC28, should advance the presence of Black prostate cancer cell lines thereby addressing prostate cancer disparity.
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DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
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作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
前列腺癌的种族差异：分子视角。
DOI：
10.2741/4515
发表时间：
2017-01-01
期刊：
Frontiers in bioscience (Landmark edition)
影响因子：
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作者：
Bhardwaj A;Srivastava SK;Khan MA;Prajapati VK;Singh S;Carter JE;Singh AP
通讯作者：
Singh AP
影响因子：
64.5
作者：
Cancer Genome Atlas Research Network
通讯作者：
Cancer Genome Atlas Research Network
影响因子：
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作者：
GAZDAR, AF;OIE, HK
通讯作者：
OIE, HK
使用下一代 DNA 测序数据进行变异发现和基因分型的框架。
影响因子：
30.8
作者：
通讯作者：
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