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Abstract Objective This study aimed to investigate the molecular mechanism of miR-150-5p regulating the malignant biological behavior of Human Epidermoid cancer cell (HEp-2) by targeting peptidyl-prolyl cis/trans isomerase NIMA-Interacting-1 (PIN1). Methods Firstly, qRT-PCR and Western blot were adopted to detect the expression levels of miR-150-5p and PIN1 in cancer tissue and paracancerous tissues of patients with LSCC, and those in human bronchial epithelial cells (16 HBE) and HEp-2. Next, the targeted relationship between miR-150-5p and PIN1 was assessed by bioinformatics website and dual-luciferase reporter assay, followed by their correlation analysis. Besides, after interfering with miR-150-5p or PIN1 expression in HEp-2 cells, CCK-8, cell colony formation assay, and transwell assay were utilized to detect the proliferation, viability, and invasion of cells, respectively. Subsequently, the protein levels of MMP-2, MMP-9, and EMT-related proteins in HEp-2 cells were checked by Western blot. Results Expression of miR-150-5p was down-regulated in LSCC tissues and HEp-2 cells. Moreover, miR-150-5p suppressed proliferation and invasion of HEp-2 cells, affected protein expression related to MMP and EMT, thereby inhibiting development of cancer. The expression of PIN1 was significantly increased in cancer tissues and HEp-2 cells, and there was a targeted relationship and negative correlation between miR-150-5p and PIN1 in cancer tissue. However, overexpression of PIN1 could reverse the effect of miR-150-5p on the proliferation and invasion of HEp-2 cells. Conclusion In a nutshell, there is a targeted relationship between PIN1 and miR-150-5p. Besides, miR-150-5p can inhibit the proliferation and invasion of HEp-2 cells by regulating the expression of PIN1. Level of evidence 3.
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Objective To investigate the expression of Pin1 protein in HepG2 cells under endoplasmic reticulum stress (ERS) and its effect on cell proliferation and apoptosis. Methods The THLE3 cells were treated with tunicamycin (TM) (TM group) or DMSO (DMSO group) for 48h. The HepG2 cells were treated with TM (TM group), ATRA(ATRA group), TM+ATRA (TM+ATRA group) or DMSO (DMSO group) for 48h. The protein levels of Bip and Pin1 were detected by Western blot, cell proliferation was detected by CCK-8 assay, and cell apoptosis was detected by flow cytometry. Results The expression of Bip both increased in THLE3 and HepG2 cells treated with TM, indicated that TM effectively induced ERS in cells. Compared with the DMSO group, the protein level of Pin1 in THLE3 cells in TM group was decreased with the increasing of TM concentration (P < 0.001). In TM+ATRA group, with the increasing of TM concentration, the expression of Pin1 was decreased in HepG2 cells (P < 0.01). The inhibitory rates was (29.33±4.73)% in HepG2 cells in TM group, significantly lower than (60.33±2.08)% in THLE3 cells in TM group (P < 0.001). In TM+ATRA group, the growth inhibition rates was increased to (60.33±6.03)% in HepG2 cells. The apoptosis rate of THLE3 cells in TM group was (22.25±0.78)%, significantly higher than (3.57±0.31)% in DMSO group (P < 0.01). The apoptosis rates of HepG2 cells in ATRA group and TM+ATRA group were significantly higher than that in the DMSO group ((10.03±0.49)% vs. (5.10±1.00)%, P < 0.05 and (23.70±0.75)% vs. (5.10±1.00)%, P < 0.01). Conclusions HepG2 cells resist ERS-induced apoptosis by maintaining Pin1 protein level. Reducing the protein level of Pin1 can significantly inhibit the cell proliferation and induce apoptosis.
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Objective:To explore the molecules mechanism of Pin1 in severe heat stroke induced acute lung injury by observing Pin1 regulate oxidative stress and apoptosis formation in pulmonary microvascular endothelial cells (PMVECs) and lung tissue in heat stressed mice.Methods:In vitro, a PMVECs heat stress (HS) model was established. In the control group, PMVECs were placed in a standard 37 °C, 5% CO 2 cell incubator; in the HS group, PMVECs were placed in a 43 °C cell incubator for 2 h, then the cells were further incubated at 37 °C for 1, 3, 6 or 12 h. PMVECs were pretreated with Pin1 inhibitor Juglone (1 μmol/L) 1 h before 43 °C of HS. In vivo, a severe heat stroke mouse model was established. In the HS group, the mice were kept at the simulation of climate chamber with temperature (35.5±0.5) °C, humidity (60±5)%, the rectum temperature in mice was measured by the anal rectal temperature table, when the temperature reached 42 °C, the heat exposure was stopped, and the mice were sacrificed at 1, 3, 6 or 12 h after HS. In the control group, the mice were kept at room temperature (25±0.5) °C. Mice received daily intraperitoneal administration of Pin1 inhibitor Juglone (1 mg/kg) for 3 d before HS. The protein level of Pin1, cleaved caspase-9 and cleaved caspase-3 were analyzed by Western blot, the level of O 2-˙ in cells was observed by DHE staining and fluorescence microscopy, the levels of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in lung tissue were measured by ELISA, the pathological changes of mice in different group were detected by HE staining, and the expression of Pin1 in the lung tissue of different groups was detected by immunohistochemical staining, the apoptosis rate of the lung tissue in different groups was tested by TUNEL staining. Results:At 1 h after HS, the protein expression of Pin1 in PMVECs and lung tissue began to increase in a rewarming time-dependent manner ( F=771.6, P<0.05; F=1 035, P<0.05). Cleaved caspase-9 protein in PMVECs and lung tissue started to increase at 3 h post-HS, then increased with a rewarming time-dependent manner ( F=729.8, P<0.05; F=1 773, P<0.05). The protein expression of cleaved caspase-3 in PMVECs and lung tissue also started to increase at 3 h after HS and the expression continued to be increased with prolonged rewarming time, and the trend was consistent with cleaved caspase-9 ( F=1 084, P<0.05; F=1 252, P<0.05). In addition, HS induced the increased release of O 2-˙ from PMVECs, HS induced the imbalance of oxidation-antioxidant system in lung tissue of mice after HS which verified by the continuous release of MDA ( F=114.2, P<0.05) and the continuous inhibition of SOD activity ( F=99.15, P<0.05). Compared with the HS group, pretreatment with Pin1 inhibitor Juglone in PMVECs and mice before HS significantly inhibited the protein expression of Pin1, cleaved caspase-9 and cleaved caspase-3 (all P<0.05), pretreatment with Pin1 inhibitor greatly reduced the release of O 2-˙ in PMVECs after HS, and promoted the restore of the oxidation-antioxidant system balance of lung tissue in mice with severe heat stroke. In addition, compared with the HS group, inhibiting the expression of Pin1 significantly decreased HS induced MDA release [(11.53±0.84) nmol/mL vs (9.65±0.69) nmol/mL, t=12.52, P<0.05], promoted the restore of SOD activity [(41.18±3.45) U/mL vs (57.52±4.83) U/mL, t=5.57, P<0.05] and improved the pathological damage of lung tissue as well as decreased the occurrence of apoptosis in post-HS mice. Conclusion:It was confirmed that Pin1 is involved in heat stress induced acute lung injury mainly through mediating oxidative stress response and apoptosis.
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Peptidyl-prolyl cis-trans isomerase Pin1 is over-expressed in prostate cancer cells and the level of expression correlates with the malignancy grade and prognosis in patients. In this work, twenty-one 2-(1H-benzimidazol-2-ylthio) acetic acid derivatives were designed and prepared with the aid of the crystal structure of Pin1 and our previous work. The chemical structures of the target compounds were confirmed by 1H NMR, 13C NMR, ESI-MS and IR. The inhibitory activity of compounds 6a-6i and 13a-13i against Pin1 were determined using a protease-coupled assay. The results indicated that twenty compounds were significantly superior to the positive control drug Juglone, and 6g, 6h and 13i exhibited the most potent Pin1 inhibitory activity, with IC50 values at the sub-micromolar level. The in vitro anti-proliferative activities of these analogs were evaluated by the MTT assay and several showed a moderate effect in human prostate cancer PC-3 cells. Molecular docking studies demonstrated that both the benzimidazole skeleton and the thioacetic acid fragment were indispensable for the compounds to interact with key residues in the catalytic domain of Pin1.
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Objective:Pin1 plays an important role in the pathogenesis of cardiovascular disease,our study aims to investigate the effects of Pin1 silencing by siRNA on H9c2 apoptosis induced by hypoxia/reoxygenation.Methods:H9c2 cells were cultured and subjected to a hypoxia/reoxygenation (H/R) condition in vitro,mimicking ischemic/reperfusion injury in vivo.The mRNA and protein expression of Pin1 were detected by RT-qPCR and Western blot.H9c2 cells were divided into control group,H/R group,H/R+Pin1 siRNA group,H/R+scramble siRNA group.MTT and flow cytometry with Annexin V-FITC/PI staining were respectively performed to detect cell viability and apoptosis.The expression of Bax and Bcl-2 were measured by Western blot.The activity of Caspase-3 was detected by automatic biochemistry analytic instrument.Results:The mRNA and protein levels of Pin1 were highly expressed in the cells of H/R group.Transfection with Pin1 siRNA strikingly inhibited the expression of Pin1.Compared with H/R group,Pin1 siRNA markedly increased cell viability,decreased the cell apoptosis and the Caspase-3 activity.Furthermore,the increased Bcl-2,decreased Bax and the ratio of Bcl-2 to Bax were observed in Pin1 siRNA group (P<0.05) compared with H/R group.Conclusion:Downregulation of Pin1 protects hypoxia/reoxygenation-injured H9c2 cells from apoptosis,which is possibly through the upregulation of Bcl-2 and downregulation of Bax and Caspase-3 activity.
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Objective To investigate the relationship between Pin1 and CyclinD1 expression and the development of gastrointestinal stromal tu?mor(GIST). Methods The protein and mRNA expression of Pin1 and CyclinD1 in 85 samples of GIST and adjacent non?cancerous tissues were detected by immunohistochemistry and real?time quantitative polymerase chain reaction. Results The expression rate of Pin1 protein in GIST tis?sues(64.7%;55/85)was higher than that in adjacent non?cancerous tissues(26.7%;4/15). Similarly,the expression rate of CyclinD1 protein in GIST tissues(42.3%;36/85)was higher than that in adjacent non?cancerous tissues(6.7%;1/15). The expression of Pin1 and CyclinD1 mRNA in GIST tissues was 7.03 and 5.53 times that in adjacent non?cancerous tissues ,respectively. There was no obvious correlation between the expres?sion of Pin1 and clinicopathological parameters. The expression of CyclinD1 was positively correlated with the grade of NIH and tumor diameter (P<0.05). There was a significant correlation between the expression of Pin1 and CyclinD1 in GIST tissues. Conclusion The expression of both Pin1 and CyclinD1 was up?regulated in GIST tissues. The significant correlation between the expression of Pin1 and CyclinD1 in GIST tissues sug?gests that their synergistic effect promotes carcinogenesis and the development of GIST.
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Objective To observe the Pin1 expression pattern in skin and to establish an inducible skin specific Pin1 overexpression mouse model. Methods The mouse Pin1 gene was cloned into modified vector pTRE2 with C?terminal Myc tag. The linearized pTRE2?Pin1 DNA was micro?injected into one?cell embryos followed by implantation into foster mice to produce TRE?Pin1 transgenic mice. Results TRE?Pin1 transgenic founder mice were successfully created. These mice were crossed with transgenic tool mice K14?rtTA to create epithelial specific double transgenic progenies. Pin1 gene was induced by incorporating doxycycline into drinking water of the mice. Pin1 protein overexpression in the skin was con?firmed by Western blot and immunohistochemistry. The endogenous Pin1 protein was predominantly expressed in epidermal cells in the skin. Conclusions The inducible skin specific Pin1 overexpression mouse model is successfully established which may serve as a useful model for further study of Pin1 functions in the skin.
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Objective To explore the expression of PIN1 and Cyclin E in gastric carcinoma tissue after radical gastrectomy and its significance.Methods 46 patients received radical gastrectomy were selected as study subjects.Cancer tissues were harvested as the gastric cancer group,and the adjacent normal tissues of tumor tissue were harvested as the control group.The expression of PIN1 and Cyclin E was observed by immunohistochemistry,and the relationship with tumor infiltration,differentiation,lymphatic and distant metastasis were analyzed.A five year followup study was also applied.Results The expression of PIN1 (3 949.66 ± 157.40) and Cyclin E (5 443.07 ±240.15) was significantly higher than those of the control group[(247.67 ± 75.63),(354.92 ± 93.88)] (all P ＜0.05),and the expression of PIN1 was related with tumor differentiation,lymphatic and distant metastasis,and the expression of Cyclin E was closely related with tumor infiltration and differentiation.The 5-year survival rate of patients with the simultaneously increased expression of PIN1 and Cyclin E was 50.0％,which was significantly lower than the expression of the two separate elevated or not elevated.Conclusion The expression of PIN1 and Cyclin E in cancer tissues after radical gastrectomy is high,and could better reflect the pathological parameters and prognosis.
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Objective To investigate the expression of Pin1 in human pancreatic carcinoma as well as adjacent tissues and to discuss the role of Pinl in oncogenesis of pancreatic carcinoma. Methods Specimen of pancreatic carcinoma tissues and adjacent tissues were collected from 20 cases. Pin1 mRNA and protein expression in pancreatic neoplasm and corresponding adjacent nontumorous tissues were detected by real-time quantitative reverse transcription-polymerase chain reaction (RQ-RT-PCR) and western blot. Results Pin1 was overexpressed at mRNA and protein level in pancreatic carcinoma tissues compared with that in their nontumorous counterparts ( 2.78 ± 1.02 vs 4.36 ± 1.27;5. 48 ± 1.69 vs 9.97 ± 1.86, P ＜ 0.05 ). Pin1 expression was not correlated to clinical stage and pathological grading of the carcinoma. Conclusion Pin1 overexpression may play a key role in pancreatic carcinoma.
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In the recent years, knowledge about cancer biomarkers has increased tremendously providing great opportunities for improving the management of cancer patients by enhancing the efficiency of detection and efficacy of treatment. Recent technological advancement has enabled the examination of many potential biomarkers and renewed interest in developing new biomarkers. Biomarkers of cancer could include a broad range of biochemical entities, such as nucleic acids, proteins, sugars, lipids, and small metabolites, cytogenetic and cytokinetic parameters as well as whole tumour cells found in the body fluid. A comprehensive understanding of the relevance of each biomarker will be very important not only for diagnosing the disease reliably, but also help in the choice of multiple therapeutic alternatives currently available that is likely to benefit the patients. This review provides a brief account on various biomarkers for diagnosis, prognosis and therapeutic purposes, which include markers already in clinical practice as well as various upcoming biomarkers.
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Antigens, Neoplasm/diagnosis , DNA, Viral/diagnosis , Epigenesis, Genetic/genetics , Hepatitis B Surface Antigens/diagnosis , Humans , Neoplasms/diagnosis , Neoplasms/therapy , Neoplastic Cells, Circulating , Neoplastic Stem Cells/cytology , T-Lymphocytes, Regulatory/cytology , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolismABSTRACT
Objective:To study the expression and relationship of Pin1 and CyclinD1 in adult papilloma of larynx,and the effect of both in laryngeal papilloma'S canceration.Method:Ninety-two cases of paraffin section with immunoperoxidase(SP)staining method was used to detect the distribution of Pin1 and CyclinD1 in 10 cases of laryngeal normal epithelial tissue,39 cases of laryngeal papilloma,27 cases of laryngeal papilloma with middle,severe atypical hyperplasia and 16 cases of laryngeal carcinoma.Result:The distribution of Pin1 and CyclinD1 increased gradually from laryngeal normal epithelial tissue tO laryngeal carcinoma(P＜0.05);No difference of the expression of CyclinD1(not including Pin1,for Pin1,P=0.009)was found between laryngeal papilloma and laryngeal papilloma with middle,severe atypical hyperplasia(P＞0.0125),but there had significant difference of the expression of Pin1 and CyclinD1 among the rest groups;There was significantly direct correlation between the expression of Pin1 and CyelinD1(P＜0.05).Conclusion:The hyper-expressions of Pin1 and CyclinD1 may play a key role in laryngea1 papilloma's malignant change.Pin1 up-regulating the expressions of cyclinD1 possibly participate in its malignant change.
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In order to investigate the expression levels of Pin1 mRNA and protein in cervical cancer and its association with Ki67 and their clinical significance, amplification of Pin1 gene was examined by RT-PCR, and the expression of both Pin1 and Ki67 protein was detected by immunohistochemistry in cervical cancer tissues. It was shown that the expression levels of Pin1 were higher in cervical cancer than in normal cervical tissues (P＜0.05). The expression of Pin1 protein was increased progressively along with the disease process from normal cervix to CIN and to cervical cancer (P＜0. 05). No significant difference in the Pin1 expression was found between disease stages (FIGO),pathological grades or pelvic lymph node metastasis status (P＞0.05). The expression of Pin1 was significantly higher in adenocarcinoma than insquamous carcinoma of the uterine cervix (P＜0.05).In cervical cancer, the overexpression of Pin1 was positively correlated with that of Ki67 (P＜0.05). These results suggested that the overexpression of Pin1 was closely related with cancer cell proliferation or progression of cervical cancer and contributed to oncogenesis. Pin1 may serve as a potential marker for cervical cancer diagnosis.
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The peptidyl prolyl cis/trans isomerase Pin1 specifically binds phosphorylated Ser/Thr-Pro protein motifs and catalyzes the cis/trans isomerization of the peptide bond,changes the conformation and influences the stability and activity of the substrates.To date,a subset of proteins has been identified as substrates for Pin1.Pin1 interacts with its substrates and plays crucial roles in cell cycle,neural pathology and immune response.Accumulating studies have revealed that Pin1 isomerase activity is regulated by its post-translational modifications,including phosphorylation and oxidation.Over-expression or regulative imbalance of Pin1 plays an important role in the pathogenesis and therapeutics of human diseases such as cancer and AD.
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Objective To investigate the expression of Pin1 and cyclin D1 in human pancreatic carcinoma and to discuss its role in oncogenesis of pancreatic cancer.Methods The mRNA expression of Pin1 and cyclinD1 in pancreatic tumor tissues and corresponding adjacent nontumor tissues 27 patients was detected by the real-time quantitative reverse transcription-polymerase chain reaction(RQ-RT-PCR).The results of RQ-RT-PCR were analyzed by one-way ANOVA and Fisher's exact probabilities test.Results Cyclin D1 and Pin1 were overexpressed at mRNA level in pancreatic carcinoma tissues compared with their adjacent nontumor tissues.Cyclin D1 overexpression were found in 14 of 20 pancreatic carcinoma tissue specimens and Pin1 overexpression in 13 of 20 carcinoma tissue specimens.The expression of cyclin D1 and Pin1 in pancreatic cystadenoma tissues was not different than that of corresponding adjacent nontumor pancreatic tissue.Pin1 overexpression positively correlated with an increase in cyclin D1 levels as shown by Fisher's exact probabilities test.However,Pin1 and cyclin D1 expression was not correlated with clinical stage and pathological parameters.Conclusions The overexpression of Pin1 in pancreatic carcinoma tissues could promote cyclin D1 expression,which might be a critical event in oncogenesis of pancreatic carcinoma.Pin1 may play a key role in pancreatic carcinoma.