[Infiltrating mast cells promote neuroendocrine differentiation and increase docetaxel resistance of prostate cancer cells by up-regulating p21]

Infiltrating mast cells up-regulates p21 to promote NED and increase docetaxel resistance in PCa cells in vitro. 目的: 探索肥大细胞浸润对前列腺癌细胞神经内分泌分化(NED)及多西他赛化疗敏感性的影响。 方法: 利用Transwell小室将人前列腺癌细胞系(LNCaP和C4-2)与人肥大细胞系(HMC-1)进行共培养，通过sh-AR慢病毒沉默C4-2细胞AR的表达，通过sh-p21质粒(pLKO.1-sh-p21)和p21过表达质粒(pCMV-p21)转染分别沉默和过表达前列腺癌细胞p21，倒置显微镜观察LNCaP和C4-2细胞的形态变化，MTT实验及克隆形成实验检测LNCaP和C4-2的细胞增殖能力，CCK8实验检测C4-2在多西他赛化疗下的细胞活性，RT-qPCR实验及Western blot实验分别检测前列腺癌细胞细胞神经内分泌标志物、AR、p21的mRNA及蛋白表达量。 结果: HMC-1细胞共培养可以增加LNCaP和C4-2细胞的NE表型表达，抑制LNCaP和C4-2细胞的增殖能力，并上调p21的表达(P < 0.05)。P21通过不依赖AR的信号通路正向调控前列腺癌细胞NED，敲除p21表达可部分逆转HMC-1共培养促进NED的作用。同时，肥大细胞共培养后的前列腺癌细胞对多西他赛化疗的抵抗性增加(P < 0.05)，敲除p21部分逆转了肥大细胞提高多西他赛化疗抵抗性的作用。 结论: 肥大细胞可以部分通过上调p21表达促进前列腺癌细胞NED以及增加对多西他赛化疗的抵抗性。

Human PCa cell lines (LNCaP and C4-2) were co-cultured with human mast cell line (HMC-1) in Transwell chambers. Androgen receptor (AR) was silenced in C4-2 cells using sh-AR lentivirus, and p21 was knocked down and overexpressed by transfecting C4-2 cells with pLKO.1-sh-p21 and pCMV-p21, respectively. The morphological changes of LNCaP and C4-2 cells were observed. MTT assay and colony formation assay were used to assess the proliferation of LNCaP and C4-2 cells. CCK8 assay was used to detect the cell viability of C4-2 cells following docetaxel trreatment. RT-qPCR and Western blotting were performed to determine the mRNA and protein expressions of neuroendocrine markers, AR and p21 in the cells.

To investigate the effect of infiltrating mast cells on neuroendocrine differentiation (NED) and docetaxel sensitivity of prostate cancer (PCa) cells in vitro.

Co-culture with HMC-1 cells enhanced the neuroendocrine phenotypes, inhibited the proliferation and up-regulated the expression of p21 in LNCaP and C4-2 cells. P21 positively regulated NED through a non-AR-dependent signaling pathway, while p21 knockdown partially reversed NED promoted by the mast cells. PCa cells co-cultured with HMC-1 cells showed increased resistance to docetaxel, and silencing p21 partially reversed docetaxel resistance in PCa cells.