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目的干涉前列腺癌细胞系VCaP细胞的CD326的表达,观察其对雄激素受体(AR)、前列腺特异性抗原(PSA)的表达以及前列腺癌细胞表型的影响,明确CD326与雄激素抵抗性前列腺癌发生之间的相关性。方法构建CD326短发夹RNA(CD326shRNA)稳定感染的VCaP细胞株。Western blot法检测稳定感染细胞株中AR及PSA的水平变化,ELISA检测VCaP细胞培养上清中的PSA分泌水平。MTT法检测细胞增殖、末端脱氧核苷酸转移酶介导的d UTP缺口末端标记法(TUNEL)和Alexa Fluor488标记的膜联素Ⅴ/碘化丙锭(annexinⅤ-Alexa Fluor488/PI)双染法检测细胞凋亡、细胞划痕实验检测细胞迁移、TranswellTM实验检测细胞侵袭能力。结果下调VCaP前列腺癌细胞株中CD326表达后,VCaP细胞中AR及PSA的表达水平显著降低,细胞上清中PSA的分泌水平也显著降低,同时伴随细胞增殖、抗凋亡、侵袭以及迁移能力的显著降低。结论下调VCaP细胞CD326水平可降低AR的表达,使前列腺癌细胞增殖、侵袭和迁移能力降低。
Objective To investigate the effect of CD326 on the expression of androgen receptor (AR), prostate-specific antigen (PSA) and prostate cancer cell line VCaP in prostate cancer cell line VCaP, and to determine the relationship between CD326 and androgen-resistant prostate Correlation between carcinogenesis. Methods The VCaP cell line stably infected with CD326 short hairpin RNA (CD326 shRNA) was constructed. The level of AR and PSA in stable infected cell lines was detected by Western blot and the level of PSA secretion in the culture supernatant of VCaP cells was detected by ELISA. MTT assay was used to detect cell proliferation, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and Alexa Fluor488 labeled annexin V-Alexa Fluor488 / PI ) Double staining method to detect apoptosis, cell scratch test to detect cell migration, TranswellTM test cell invasive ability. Results The down-regulation of CD326 expression in VCaP prostate cancer cell lines significantly reduced the expression of AR and PSA in VCaP cells, and significantly decreased the secretion of PSA in the supernatant of the VCaP cells, accompanied by cell proliferation, anti-apoptosis, invasion and migration Significantly lower. Conclusion The downregulation of CD326 in VCaP cells can decrease the expression of AR and decrease the proliferation, invasion and migration of prostate cancer cells.