Zhao Jiafu, Ma Xiaoyan, Xu Houqiang
Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, Guizhou University, Guiyang, Guizhou 550025, P.R. China.
College of Animal Science, Guizhou University, Guiyang, Guizhou 550025, P.R. China.
Oncol Lett. 2022 Jul 1;24(2):289. doi: 10.3892/ol.2022.13409. eCollection 2022 Aug.
Prostate cancer (PCa) is one of the most common malignant tumours in the world and seriously affects health of men. Studies have shown that microRNA (miR)-29b-3p and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein epsilon (YWHAE) play important roles in influencing the proliferation and apoptosis of PCa cells. However, the molecular mechanism of miR-29b-3p and YWHAE in the proliferation and apoptosis of PCa cells remains unclear. In the present study, bioinformatics as well as and experiments were used to predict and verify the targeting relationship between YWHAE and mir-29B-3p and investigate the potential roles of YWHAE and mir-29b-3p in the proliferation and apoptosis of 22RV1 cells. Using bioinformatics and a double luciferase system assay, it was confirmed that miR-29b-3p can target YWHAE 3'untranslated region and affect the expression of YWHAE, suggesting that miR-29b-3p may be a potential miRNA of YWHAE. Reverse transcription-quantitative PCR, Cell Counting Kit-8, Transwell and cell scratch assays showed that miR-29b-3p significantly inhibited the proliferation, invasion and migration of 22Rv1 cells (P<0.01). Rescue experiments demonstrated that YWHAE gene introduction reversed the inhibitory effect of miR-29b-3p on 22Rv1 cells. Western blotting revealed that the upregulation of miR-29b-3p inhibited YWHAE expression, resulting in a very significant decrease in the ratio of p-BAD/BAD and full-length caspase 3/cleaved caspase 3 (P<0.01) and an extremely significant increase in the ratio of BAX/BCL-2 (P<0.01). A tumourigenesis test in nude mice confirmed that the upregulation of miR-29b-3p inhibited tumour growth by targeting YWHAE. The present experiments confirmed that miR-29b-3p plays a tumour suppressor role in 22Rv1 PCa cells, and the YWHAE/BCL-2 regulatory axis plays a vital role in miR-29b-3p regulating the proliferation and apoptosis of 22Rv1 cells. These results may provide a theoretical basis for the diagnosis and targeted treatment of PCa.
前列腺癌(PCa)是全球最常见的恶性肿瘤之一,严重影响男性健康。研究表明,微小RNA(miR)-29b-3p和酪氨酸3-单加氧酶/色氨酸5-单加氧酶激活蛋白ε(YWHAE)在影响PCa细胞的增殖和凋亡中起重要作用。然而,miR-29b-3p和YWHAE在PCa细胞增殖和凋亡中的分子机制仍不清楚。在本研究中,利用生物信息学以及实验来预测和验证YWHAE与mir-29B-3p之间的靶向关系,并研究YWHAE和mir-29b-3p在22RV1细胞增殖和凋亡中的潜在作用。通过生物信息学和双荧光素酶系统检测,证实miR-29b-3p可靶向YWHAE的3'非翻译区并影响YWHAE的表达,提示miR-29b-3p可能是YWHAE的潜在微小RNA。逆转录定量PCR、细胞计数试剂盒-8、Transwell和细胞划痕实验表明,miR-29b-3p显著抑制22Rv1细胞的增殖、侵袭和迁移(P<0.01)。挽救实验表明,导入YWHAE基因可逆转miR-29b-3p对22Rv1细胞的抑制作用。蛋白质免疫印迹法显示,miR-29b-3p的上调抑制了YWHAE的表达,导致p-BAD/BAD和全长半胱天冬酶3/裂解的半胱天冬酶3的比值显著降低(P<0.01),BAX/BCL-2的比值极显著升高(P<0.01)。裸鼠成瘤实验证实,miR-29b-3p的上调通过靶向YWHAE抑制肿瘤生长。本实验证实miR-29b-3p在22Rv1前列腺癌细胞中起肿瘤抑制作用,YWHAE/BCL-2调控轴在miR-29b-3p调节22Rv1细胞的增殖和凋亡中起关键作用。这些结果可能为前列腺癌的诊断和靶向治疗提供理论依据。