The novel long noncoding RNA LOC283070 is involved in the transition of LNCaP cells into androgen-independent cells via its interaction with PHB2 / 亚洲男科学杂志(英文版)

We sought to investigate the underlying mechanism of action of the long noncoding RNA (lncRNA) LOC283070 in the development of androgen independence in prostate cancer. The interactions between LOC283070 and target proteins were investigated by RNA pull-down and RNA-binding protein immunoprecipitation (RIP) assays. Subcellular fractionation and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were used to detect the subcellular localization of LOC283070. Western blotting was performed to detect the expression of prohibitin 2 (PHB2). Luciferase activity assays were performed to evaluate the effects of LOC283070 and PHB2 on the androgen receptor (AR) signaling pathway. A methyl thiazolyl tetrazolium (MTT) assay and a growth curve assay were used to test cell viability. Flow cytometry was performed to analyze cell cycles. A transwell assay was employed to test cell migration. We identified PHB2 as an interaction partner of LOC283070 in the pull-down and RIP experiments. Furthermore, we confirmed that the enrichment of LOC283070 with PHB2 in androgen-independent LNCaP (LNCaP-AI) cells was much greater than that in LNCaP cells. Moreover, the expression of PHB2 was not significantly different between the two cell lines, and the expression of LOC283070 in the nuclei of the LNCaP-AI cells was significantly greater than that in the LNCaP cells. In vitro data revealed that PHB2 overexpression significantly inhibited AR activity and cell proliferation and migration and induced accumulation of prostate cancer cells in G0/G1 phase. Moreover, the overexpression of LOC283070 fully abrogated the effects of PHB2 overexpression. In conclusion, we found that LOC283070 can bind to PHB2 located in the nucleus and inhibit its effect, and this is one of the mechanisms by which LOC283070 is involved in the transition of LNCaP cells into androgen-independent cells.

The novel long noncoding RNA LOC283070 is involved in the transition of LNCaP cells into androgen-independent cells via its interaction with PHB2 / 亚洲男科学杂志(英文版)

We sought to investigate the underlying mechanism of action of the long noncoding RNA (lncRNA) LOC283070 in the development of androgen independence in prostate cancer. The interactions between LOC283070 and target proteins were investigated by RNA pull-down and RNA-binding protein immunoprecipitation (RIP) assays. Subcellular fractionation and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were used to detect the subcellular localization of LOC283070. Western blotting was performed to detect the expression of prohibitin 2 (PHB2). Luciferase activity assays were performed to evaluate the effects of LOC283070 and PHB2 on the androgen receptor (AR) signaling pathway. A methyl thiazolyl tetrazolium (MTT) assay and a growth curve assay were used to test cell viability. Flow cytometry was performed to analyze cell cycles. A transwell assay was employed to test cell migration. We identified PHB2 as an interaction partner of LOC283070 in the pull-down and RIP experiments. Furthermore, we confirmed that the enrichment of LOC283070 with PHB2 in androgen-independent LNCaP (LNCaP-AI) cells was much greater than that in LNCaP cells. Moreover, the expression of PHB2 was not significantly different between the two cell lines, and the expression of LOC283070 in the nuclei of the LNCaP-AI cells was significantly greater than that in the LNCaP cells. In vitro data revealed that PHB2 overexpression significantly inhibited AR activity and cell proliferation and migration and induced accumulation of prostate cancer cells in G0/G1 phase. Moreover, the overexpression of LOC283070 fully abrogated the effects of PHB2 overexpression. In conclusion, we found that LOC283070 can bind to PHB2 located in the nucleus and inhibit its effect, and this is one of the mechanisms by which LOC283070 is involved in the transition of LNCaP cells into androgen-independent cells.

Effect of all-trans retinoic acid on expressions of Prohibitin1 and Prohibitin2 in hypoxic damage of renal tubular epithelial cells / 中华实用儿科临床杂志

Objective To investigate the effect of all-trans retinoic acid(ATRA) on the expressions of Prohibitin1 (PHB1) and Prohi-bitin2 (PHB2) in hypoxic damage of renal tubular epithelial cells (RTEC).Methods Rat proximal tubular epithelial cell line NRK-52E culture was performed in the 37 ℃ 50 mL/L carbon dioxide incubator with mixture medium of fetal bovine serum and double-antibody(100 mL medium plus 5 mL fetal calf serum and 1 mL double-antibody).After 3 times cell propagation,the cells were divided into 3 groups:normal group,model group and ATRA intervention group.The normal group wasn't disposed,and the model group was put into vacuum tank filled with hypoxic gas(950 mL/L nitrogen and 50 mL/L carbon dioxide) to induce a hypoxic damage of RTEC.The ATRA intervention group was added 0.1 μmol/L ATRA and was treated with hypoxic gas as model group.After 24 h and 36 h,the mRNA expressions of PHB1,PHB2 and transforming growth factor-β1 (TGF-β1) were measured by using real-time PCR method,and the protein expressions of PHB1,PHB2 and TGF-β1 were detected by using Western blot method.Results 1.Compared with normal group,NRK-52E cells PHB1,PHB2 protein expressions and mRNA expressions at 2time points(24 h,36 h) were significantly decreased in model group and ATRA intervention group (all P ＜ 0.05),and the longer hypoxia time,the lower expression value.Compared with model group,NRK-52E cells PHB1 and PHB2 protein expressions and mRNA expressions in ATRA intervention group were increased significantly at 2 time points (all P ＜ O.05).2.Compared with normal group,NRK-52E cells TGF-β1 expressions and mRNA expressions at 2 time points(24 h,36 h) were significantly increased in model group and ATRA intervention group(all P ＜ 0.05),and the longer hypoxia time,the higher expression value;Compared with model group,NRK-52E cells TGF-β1 protein expressions and mRNA expressions in ATRA intervention group were decreased significantly at 2 time points(all P ＜ 0.05).Conclusions ATRA can significantly up-regulate the mRNA expressions and protein expressions of PHB1 and PHB2 in hypoxic damage of RTEC,and ATRA may have a protective effect against hypoxic damage.