Takezawa Yuta, Izumi Kouji, Machioka Kazuaki, Iwamoto Hiroaki, Naito Renato, Makino Tomoyuki, Kadomoto Suguru, Natsagdorj Ariunbold, Kadono Yoshifumi, Keller Evan T, Zhang Jian, Mizokami Atsushi
Integrative Cancer Therapy and Urology, Division of Cancer Medicine, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan.
Integrative Cancer Therapy and Urology, Division of Cancer Medicine, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
Anticancer Res. 2018 Apr;38(4):2045-2055. doi: 10.21873/anticanres.12444.
To investigate how androgen-sensitive LNCaP cells crosstalk with androgen-insensitive DU145 or PC-3 cells.
The numbers of LNCaP cells were counted when co-cultured with DU145 or PC-3 cells and vise versa. Androgen receptor (AR) activity in LNCaP cells was examined by luciferase reporter assay after transfection with a luciferase reporter driven by PSA promoter in the presence of DU145 or PC-3 cells. Concentration of androgens in the medium was measured by liquid chromatography-mass spectrometry (LC-MS/MS). The ability of migration and invasion of PC-3 and DU145 cells was investigated using a 2-layer chamber, in the presence of LNCaP cells.
Co-culture of LNCaP cells with DU145 cells resulted in the conversion of dehydroepiandrosterone (DHEA) to dihydrotestosterone (DHT), which stimulated cell proliferation and PSA promoter activity in LNCaP cells. The increased cell proliferation rate and AR activity, induced in LNCaP cells after DHT treatment, was further enhanced by co-culture with DU145 cells. LNCaP cells also stimulated the proliferation of DU145 and PC-3 cells, via secreting soluble factors. Finally, LNCaP cells promoted migration and invasion of PC-3 cells, in a co-culture system; however inhibited migration and invasion of DU145 cells.
Crosstalk between androgen-sensitive PCa cells and androgen-insensitive PCa cells might develop the progression of PCa.
研究雄激素敏感的LNCaP细胞如何与雄激素不敏感的DU145或PC-3细胞进行细胞间通讯。
将LNCaP细胞与DU145或PC-3细胞共培养时对LNCaP细胞进行计数，反之亦然。在用前列腺特异性抗原（PSA）启动子驱动的荧光素酶报告基因转染LNCaP细胞后，在存在DU145或PC-3细胞的情况下，通过荧光素酶报告基因测定法检测LNCaP细胞中的雄激素受体（AR）活性。通过液相色谱-质谱联用（LC-MS/MS）测定培养基中雄激素的浓度。在存在LNCaP细胞的情况下，使用双层小室研究PC-3和DU145细胞的迁移和侵袭能力。
LNCaP细胞与DU145细胞共培养导致脱氢表雄酮（DHEA）转化为双氢睾酮（DHT），后者刺激LNCaP细胞的细胞增殖和PSA启动子活性。DHT处理后LNCaP细胞中诱导的细胞增殖率增加和AR活性增加，通过与DU145细胞共培养进一步增强。LNCaP细胞还通过分泌可溶性因子刺激DU145和PC-3细胞的增殖。最后，在共培养系统中，LNCaP细胞促进PC-3细胞的迁移和侵袭；然而抑制DU145细胞的迁移和侵袭。
雄激素敏感的前列腺癌细胞与雄激素不敏感的前列腺癌细胞之间的细胞间通讯可能促进前列腺癌的进展。
