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Objective:To explore the effect of miR-181a on chondrosarcoma cell growth through phosphatase and tensin homolog(PTEN) and its possible regulatory mechanism.Methods:From January to December 2022, 10 fresh chondrosarcoma and 10 chondroma tissues from orthopedic patients of Hunan Provincial People′s Hospital were collected, and the expression of miR-181a in chondrosarcoma and chondroma tissues was detected using real-time fluorescence quantitative polymerase chain reaction (qRT-PCR); Chondrosarcoma cell SW1353 was cultured in vitro and transfected with miR-181a inhibitor (miR-181a inhibition group) and control (miR-NC, control group), respectively. The effects of miR-181a on the growth and proliferation of SW1353 cells were detected by cell counting kit (CCK-8) and clone formation test, respectively; The binding sites between miR-181a and PTEN were predicted through the Target Scan database, and verified using dual luciferase reporter gene experiments; The mimetic miR-181a (miR-181a group) and its control (miR-NC, control group) were transfected into chondrosarcoma cell SW1353, respectively. The adenosine triphosphate (ATP) content, glucose consumption, and lactic acid production in the cells were measured, and the effect of miR-181a on glycolysis of SW1353 cells was analyzed. Results:The expression of miR-181a in chondrosarcoma tissues was significantly higher than that in chondroma tissues ( P<0.05). The cell growth and clonogenesis ability of miR-181a inhibition group were significantly lower than those of control group (all P<0.05). It was predicted by Target Scan online website that there might be binding sites between miR-181a and PTEN, and co-transfection of wild-type PTEN and miR-181a could significantly reduce luciferase activity by double luciferase reporter assay ( P<0.05). The ATP content, glucose consumption and lactic acid production of miR-181a group were significantly higher than those of miR-NC group (all P<0.05). Conclusions:MiR-181a promotes the growth of chondrosarcoma cells through PTEN-mediated glycolysis.
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Objective:To investigate the effect of miR-26a mediated phosphatidylinositol-3 kinase (PI3K)/protein kinase B (AKT) signaling pathway on angiogenesis in rats with cerebral ischemia.Methods:A total of 100 male SD rats were divided into sham operation group, model group, miR-NC group, and miR-26a group according to the random number table method. The miR-NC group and the miR-26a group were injected with 5 μl miR-26a simulant negative control and miR-26a simulant into the lateral ventricle respectively. The sham operation group and the model group were injected with the same amount of normal saline respectively. The middle cerebral artery occlusion model was induced by the modified intraluminal suture method. In the sham operation group, the thread was only inserted without ligation. Five rats in each group were injected intraperitoneally with 5-bromodeoxyuridine (BrdU) daily for 7 days. Rat brain microvascular endothelial cells (BMECs) cultured and transfected in vitro were divided into control group, oxygen glucose deprivation (OGD) group, miR-NC group, and miR-26a group. The dual luciferase experiment verified the regulatory effect of miR-26a on the phosphatase and tensin homolog deleted on chromosome 10 (PTEN). Longa score was used to detecte the neurological damage of rats. The volume of cerebral infarction was measured by triphenyltetrazolium chloride (TTC) staining. The methyl thiazolyl tetrazolium (MTT) staining, annexin Ⅴ fluorescein isothiocyanate/propidium iodide double staining and tubule formation experiment were used to detect the proliferation, apoptosis and angiogenesis of BMECs, respectively. Real-time fluorescence quantitative reverse transcription polymerase chain reaction was used to detect the miR-26a expression of ischemic brain tissue and BMECs. Immunofluorescence double labeling method (BrdU/von Willebrand factor [vWF]) was used to detect the proliferation of rat vascular endothelial cells. Western blot analysis was used to detect the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), angiopoietin-2 (Ang-2), PTEN, PI3K and Akt protein in ischemic brain tissue.Results:Bioinformatics and dual luciferase experiments verified the targeted regulation of PTEN by miR-26a. Compared with the sham operation group, the expression of miR-26a, VEGF, bFGF, Ang-2, PI3K, AKT and the number of BrdU + /VWF + cells in ischemic brain tissue in the model group and miR-NC group increased, while the expression of PTEN decreased (all P<0.05). Compared with the model group, the effect of various indexes in the miR-26a group was more significant (all P<0.05). Compared with the control group, the proliferation and angiogenesis of BMECs in the OGD group and the miR-NC group were significantly increased, and the apoptosis was significantly reduced (all P<0.05). Compared with the OGD group, the effect of various indexes in the miR-26a group was more significant (all P<0.05). Conclusion:miR-26a can mediate the targeted inhibition of PTEN expression, up-regulate angiogenesis related factors (VEGF, bFGF and Ang-2), and promote vascular endothelial cell proliferation and angiogenesis in rats with cerebral infarction by activating PI3K/Akt signaling pathway.
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Abstract: Cutaneous metastases are uncommon in daily practice, although very important, since they may be the first manifestation of an undiscovered primary neoplasm or the first indication of recurrence. Cutaneous metastases from the breast are the most frequent in women and cutaneous metastases from the lung are the most frequent in men. Thyroid carcinoma, despite representing the most frequent endocrine neoplasm, is considered a rare neoplasm, corresponding to 1% of malignant neoplasms diagnosed. Cutaneous metastases from follicular carcinoma are rare and occur mainly in the head and neck area. We report a case of cutaneous metastasis in a patient with follicular thyroid carcinoma and breast carcinoma. Because of the association of these two neoplasms, the possibility of Cowden Syndrome - multiple hamartoma syndrome - was raised, but was excluded by genetic analysis of PTEN gene.
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Humans , Female , Middle Aged , Skin Neoplasms/secondary , Breast Neoplasms/pathology , Thyroid Neoplasms/pathology , Adenocarcinoma, Follicular/secondary , Neoplasms, Multiple Primary/pathology , Skin Neoplasms/diagnosis , Biopsy , Breast Neoplasms/diagnosis , Thyroid Neoplasms/diagnosis , Immunohistochemistry , Adenocarcinoma, Follicular/diagnosis , Neoplasms, Multiple Primary/diagnosisABSTRACT
Objective To investigate germline mutation of breast cancer susceptibility genes BRCA1/2,TP53 and PTEN in Chinese breast cancer patients. Methods All of128 female breast cancer patients in Peking University People′s Hospital from January 2016 to August 2018 were selected as objects. Among them,44 cases were sporadic breast cancer and 84 werebreast cancer patients with genetic high risks. Germline mutations of BRCA1,BRCA2,TP53 and PTENwere detected by NGS.χ2 test was used to analyze the difference of pathogenic mutation rates between sporadic breast cancer group and breast cancer with high genetic risks.Groups were divided according to the clinical features of the patients(family history, triple-negative breast cancer,age and bilateral breast cancer).Among them,there were 42 cases with family history of breast cancer,34 cases of triple-negative breast cancer,33 cases of early-onset breast cancer and 7 cases of bilateral breast cancer. Fisher′s exact probability test compared the relationship between pathogenic mutations of BRCA1/2 gene and clinical characteristics of breast cancer patients with hereditary risk factors. Results In 128 cases of breast cancer,30 germline mutations of BRCA1/2 were detected, including 13 pathogenic mutations and 3 newly discovered mutations(BRCA1:c. 4760C>G,BRCA2:c. 44134414del and BRCA2:c. 64826485del). The new mutations may be unique mutations of Chinese population. There were 3 cases of TP53 mutations,including 1 pathogenic mutation. All of the 3 mutations were found in early-onset breast cancer. Germline mutation of T53 has important detection significance for early-onset hereditary breast cancer. There were 5 cases of PTEN mutations,including 3 pathogenic mutations. Among 84 breast cancer patients with genetic high risks,the carry mutation rate was 40.5％(34/84)and the pathogenic mutation rate was 15.4(13/84). Among 44 sporadic cases,the carry mutation rate was 9％(4/44). The pathogenic mutation rate was 6.8％(3/44). Breast cancer susceptibility genes were carried at a higher rate in breast cancer patients with genetic high risks(P<0.001). BRCA1/2 mutations did not show statistical differences among groups of breast cancer patients with hereditary high risk factors . Conclusion Germline mutation detection of breast cancer susceptibility genes by next-generation sequencing is of great significance in breast cancer risk prediction and prognosis evaluation.
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Objective To study the relation between the expression of P130 Crk-associated substrate (P130Cas),phosphatase and tensin homolog (PTEN)and the epithelial-mesenchymal transition (EMT) of skin scar carcinoma.Methods Tissues of skin scar carcinoma,scar and normal skin were collected from 8 patients who were pathologically diagnosed as skin scar carcinoma with high differentiated squamous cell carcinoma.The expression of PTEN,P130Cas,E-cadherin and Vimentin in normal skin,skin scar and skin scar carcinoma tissues were detected by immunohistochemical method of S-P.Results The expression of PTEN,P130Cas and E-cadherin in normal skin,scar and skin scar carcinoma tissues were all significantly different (P ＜ 0.05).The expression of Vimentin in skin scar carcinoma tissues were significantly increased than that in normal skin and skin scar tissues,but there was no statistically significance difference between skin scar tissues and normal skin (P ＞ 0.05).The expression of PTEN in skin scar carcinoma tissues was negatively correlated with P130Cas (r =-0.78,P =0.023) and positively correlated with E-cadherin (r =0.83,P =0.011),but there were no correlation between PTEN and Vimentin (P ＞ 0.05);The expressions of P130Cas in skin scar carcinoma tissues was negatively correlated with Ecadherin (r =-0.74,P =0.035),but there were no statistically significant correlation between P130Cas and Vimentin (P＞0.05).Conclusions Both PTEN and P130Cas involved in the EMT process of skin scar carcinoma and may be an important mechanism in scar carcinogenesis.
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Objective To investigate the expressions of phosphatase and tensin homologue deleted on chromosome ten(PTEN)and epithelial cadherin(E-cadherin)in breast cancer tissues and explore their clini-cal significance. Methods We retrospectively analyzed the clinical data of 263 breast cancer patients admitted to the First Affiliated Hospital of Harbin Medical University from November 2012 to December 2014. The expressions of PTEN and E-cadherin were detected by immunohistochemistry. The relationship of protein ex-pression with clinicopathological characteristics was analyzed by χ2 test or Fisher exact test. Contingency corre-lation analysis was used to analyze the correlation between PTEN and E-cadherin,and Kaplan-Meier was used to evaluate the relationship between their expressions and patients' survival. COX regression model was used to analyze risk factors. Results The positive expression rates of PTEN and E-cadherin in breast cancer tissues were 68. 4％(180 / 263)and 95. 4％(251 / 263)respectively,lower than those in para-tumor breast tissues 77. 9％(205 / 263)and 98. 5％(259 / 263),with statistically significant differences(χ2 = 6. 056,P = 0. 014;χ2 = 4. 125,P = 0. 042). PTEN expression was associated with lymph node metastasis( χ2 = 8. 443,P =0. 015)and clinical stage(χ2 = 9. 253,P = 0. 010). E-cadherin expression was not correlated with age,meno-pausal status,tumor maximum diameter,lymph node metastasis and clinical stage(all P > 0. 05). Contingency correlation analysis showed a positive correlation between PTEN and E-cadherin expressions in breast cancer tissues(C = 0. 125,P = 0. 041). Survival analysis showed that the 5-year tumor-free survival rate was 81. 9％in the PTEN negative group,lower than 95. 0％ in the positive group(χ2 = 12. 040,P = 0. 001). The 5-year tumor-free survival rate in the E-cadherin negative group was 66. 7％ ,lower than 92. 0％ in the positive group (χ2 = 13. 313,P < 0. 001). COX multivariate analysis showed that negative expressions of PTEN and E-cadherin and lymph node metastasis were independent risk factors for the prognosis of breast cancer patients (HR = 2. 554,95％ CI:1. 016-6. 420,P = 0. 046;HR = 3. 573,95％ CI:1. 136-11. 239,P = 0. 029;HR =3. 622,95％ CI:2. 026-6. 476,P < 0. 001). Conclusion PTEN is related to E-cadherin expression and low expressions of both may be one of the mechanisms of breast cancer development,invasion and metastasis. Com-bined detection can be used as an indicator to determine the prognosis of breast cancer.
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Objective To investigate the correlation between tumor necrosis factor-α inducing protein (Tipα),phosphatase and tensin homologue deleted onchromosome ten (PTEN) and gastric cancer.Methods To detecte the relative quantification of Tipα and lost phosphatase gene of tenth chromosome PTEN in gastric cancer,paracancerous tissues and normal gastric tissues by real-time quantitative polymerase chain reaction (RT-qPCR) technique.Results (1) The expression levels of Tipα and PTEN in gastric cancer,paracancerous tissues and normal gastric tissues were 0.83 ± 0.07,0.16 ± 0.10,0.10 ± 0.12,0.23 ±0.05,0.92 ±0.07,1.84 ±0.13,respectively.Comparison of gastric cancer with paracancerous tissues and normal gastric tissue,there were significant statistical differences(P ＜0.05).(2) The expression levels of Tipα and PTEN in low and well differentiated gastric carcinoma were1.10 ± 0.04,0.36 ± 0.05,0.08 ± 0.05,0.36 ± 0.04,respectively,both of which are significantly associated with the degree of differentiation of gastric cancer(P ＜ 0.05);The expression levels of Tipα and PTEN in gastric cancer tissues with or without lymph node metastasis were 0.18 ±0.12,0.82 ±0.09,0.10 ±0.12,0.38 ±0.10,both had significant correlation with lymph node metastasis of gastric cancer(P ＜ 0.05);The expression of PTEN in gastric cancer tumor node metastasis (TNM) Ⅰ-Ⅱ and Ⅲ-Ⅳ were 0.25 ±0.04,0.06±0.07,which means PTEN was significantly correlated with gastric cancer TNM staging(P ＜ 0.05).(3) The expressions of Tipα and PTEN were negatively correlated in gastric cancer tissues (r =-0.883,P ＜ 0.05).Conclusions (1) Tipα may be a cancer-promoting factor;PTEN maybe a tumor suppressor factor;(2) There is a negative correlation between the expression of Tipα and PTEN in gastric carcinoma.
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Objective To investigate the expression of miR-29a in serum of acute kidney injury patients, and whether miR-29a can affect the apoptosis of renal tubular cells by regulating the expressions of phosphatase and tensin homologue deleted on chromosome ten (PTEN).Methods Blood samples were collected from 113 patients with acute kidney injury (AKI) and 110 healthy controls.The expressions of miR-29a in serum were detected by real-fluorescence quantitative polymerase chain reaction (RT-q-PCR).HK-2 cells were cultured in vitro, miR-29a NC or miR-29a mimic was transfected into HK-2 cells, and apoptosis of HK-2 cells was induced by transforming growth factor-β1 (TGF-β1).Flow cytometry was used to investigate the apoptosis of HK-2 cells.Bioinformatics was used to predict potential target genes of miR-29a.Luciferase reporter gene test was used to verify the complementary pairing relationship between miR-29a and PTEN.The expression of PTEN was detected by Western blot.After transfection of plasmids overexpressing PTEN, the apoptosis of HK-2 cells was detected by flow cytometry.Results Compared to the healthy control group, the serum expression of miR-29a was decreased in AKI patients (P ＜ 0.05).Flow cytometry results showed that transfection of miR-29a mimic was significantly decreased HK-2 cells apoptosis compared to miR-29 NC group (P ＜ 0.05).The Luciferase reporter assay results showed that miR-29a and PTEN had complementary relationship.After transfected with overexpression of PTEN, the HK-2 cells apoptosis rate was significantly increased (P ＜ 0.05).Conclusions In the serum of AKI patients, the expression of miR-29a was decreased, overexpression of miR-29a inhibited the apoptosis of renal tubular epithelial cells by inhibiting the expression of PTEN protein.
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Objective@#To elucidate the expression levels of key immune biomarkers, phosphate and tension homology deleted on chromosome ten (PTEN) and programmed cell death protein1(PD-1),of different immune tolerance pathway in classic Hodgkin’s lymphoma (CHL) to further determine their clinical role and prognostic significance.@*Methods@#The clinical features and prognostic factors of 56 CHL patients, who were admitted to the TianJin Medical University Cancer Institute from February 2003 to August 2013, were retrospectively analyzed. PTEN and PD-1 protein expression levels were analyzed by immunohistochemistry, Epstein-Barr virus encoded RNA (EBER) was performed by in situ hybridization assay. Correlations between the expression of biomarkers and clinicopathologic parameters were examined and survival analyses were performed.@*Results@#This cohort of 56 CHL patients included 34 males and 22 females with a median age of 25 years (ranged from 7 to 71 years). In a univariate analysis, age≥45, IPS score >2, EBER positive, high expression of PTEN protein conferred inferior 5-year OS and 5-year PFS; In a multivariate model, age≥45, IPS score >2, EBER positive, high expression of PTEN protein were identified as the independent adverse prognostic factors for CHL.@*Conclusions@#This study suggested for the first time that PTEN was independent prognostic immune biomarkers in CHL, which provided the novel therapeutic strategy of immune therapy for CHL.
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Objective To evaluate the effect of over-expression of phosphatase and tensin homolog does on chromosome ten (PTEN) in podocytes on kidney under high fat diet (HFD) in vivo and clarify the mechanism how PTEN regulates scavenger receptor A (SR-A) expression exposed to oxidized low density lipoprotein (ox-LDL) in podocytes in vitro.Methods The podocyte-specific PTEN knockin (PPKI) mice were fed with HFD to establish mouse model of lipid-induced renal injury.Mice were divided into four groups:ND+Ctrl group,ND+PPKI group,HFD+Ctrl group and HFD+PPKI group.After 24 weeks of dietary intervention,all mice were tested for clinical and biochemical parameters,including serum creatinine (Scr) as well as urine albumin excretion rate (UAER);renal lipid content was measured by oil red O staining and cholesterol quantitative analysis;the pathological changes of glomeruli were observed by PAS staining and electron microscope.Podocyte injury was induced by ox-LDL in vitro.Western blotting was used to detect the changes of SR-A expression induced by ox-LDL after YAP-siRNA interfering (si-YAP),as well as YAP phosphorylation induced by ox-LDL after interfering by PTEN-siRNA (si-PTEN) and PTEN phosphatase inhibitor (Bpv-PTEN),and overexpressing by recombinant adenovirus (ad-PTEN).Results Compared with ND+Ctrl group,HFD+ Ctrl group significantly aggravated the levels of Scr and UAER,the expression of SR-A in podocytes,renal lipid content,mesangial matrix expansion,effacement of podocyte foot processes,and incrassation of glomerular basement membrane (all P ＜ 0.05).Conversely,compared with HFD+Ctrl group,HFD+ PPKI group obviously alleviated the above lipid-induced renal damage (all P ＜ 0.05).In vitro,the expression of SR-A in podocytes was up-regulated when stimulated with ox-LDL (P ＜ 0.05),and the knockout of YAP significantly down-regulated the expression of SR-A induced by ox-LDL (P ＜ 0.05).Exposed to ox-LDL,the expression of p-YAP increased in podocytes (P ＜ 0.05);over-expression of PTEN inhibited p-YAP up-regulation induced by ox-LDL (P ＜ 0.05),while either knockdown of PTEN or inhibition of PTEN phosphatase activity displayed opposite effect (all P ＜ 0.05).Conclusions Over-expression of PTEN in podocytes protected the kidney against damage from HFD in vivo and PTEN might suppress SR-A mediated lipid uptake via dephosphorylating p-YAP to prevent podocyte injury from ox-LDL.
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Objective To investigate the expressions and significances of phosphatase and tension homologous genes (PTEN) and human telomerase reverse transcriptase (hTERT) in esophageal cancer.Methods 84 specimens of esophageal cancer underwent surgical resection from December 2010 to December 2012 were collected,at the same time,30 cases of para cancerous tissues and 30 normal esophageal mucosa tissues were taken as control.Streptavidin peroxidase (SP) immune-histochemical method was used to detect the expressions of PTEN and hTERT in the esophageal tissue specimens,according to the clinical data,the relationships between PTEN,hTERT in the esophageal cancer tissues with clinical pathological features and prognosis was analyzed.Results (1) The positive expression rate of PTEN in esophageal cancer tissue was significantly lower than that in para cancerous tissue (x2 =5.077,P =0.024) and normal mucosa tissue (x2 =22.810,P =0.000);the positive expression rate of PTEN in para cancer tissues was significantly lower than that in normal mucosa (x2 =5.104,P =0.024).(2) The positive expression rate of hTERT in esophageal cancer tissues was significantly higher than that in para cancerous tissue (x2 =19.724,P =0.000) and normal mucosa tissue (x2 =46.392,P =0.000);The positive expression rate of hTERT in para cancerous tissues was significantly higher than that in normal mucosa (x2 =4.565,P =0.033).(3) The expression intensities of PTEN and hTERT in esophageal cancer tissues were related to the degree of tumor invasion,clinical stage,lymph node metastasis and degree of differentiation (P ＜ 0.05).(4) At the end of the follow-up,the median survival period of all patients was 22 months,the cumulative survival rates of 1,3,and 5 years after operation were 65.48％,38.10％ and 28.57％ respectively.The cumulative survival rates of 1,3,and 5 years after operation in patients with low expression of PTEN and patients with high expression of PTEN were 60.00％ and 84.21％,30.77％ and 63.16％,23.08％ and 47.37％,respectively.The survival rate of patients with high expression of PTEN was significantly higher than that of patients with low expression of PTEN (x2 =5.073,P =0.024).The cumulative survival rates of 1,3,and 5 years after operation in patients with low expression of hTERT and patients with high expression of hTERT were 76.47％ and 58.00％,50.00％ and 30.00％,38.24％ and 22.00％,respectively.The survival rate of patients with high expression of hTERT was significantly lower than that of patients with low expression of hTERT (x2 =4.174,P =0.041).Conclusions The expression of PTEN was low in esophageal cancer tissues and high in hTERT,which could be used as a predictor of clinical prognosis in patients with esophageal cancer.
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Objective To explore the influence of up-regulated PTEN gene expression on apoptosis and chemosensitivity of ovarian cancer cells.Methods HEY cells transfected by exogenous PTEN plasmid (PTEN-HEY) were used as the PTEN-HEY group,and those transfected by empty vectors (pcDNA-HEY) and non transfected cells (HEY) were used as the control groups.The expressions of PTEN protein and mRNA in each group were detected with Western blot and quantitative real-time polymerase chain reaction (qRT-PCR),the influence of PTEN overexpression on HEY cells apoptosis was evaluated with flow cytometry,and tumorigenicity in vivo was evaluated with nude mouse model tumor formation test,and cisplatin sensitivity of tumor cells was evaluated with methyl thiazolyl tetrazolium (MTT) detection in condition of PTEN overexpression.Results Compared to those in the pcDNA-HEY group and HEY group,PTEN protein and mRNA expression levels significantly increased after PTEN gene transfected (PTEN-HEY group) in 24 h.Compared to those in the pcDNA-HEY group and HEY group,the apoptosis rate of PTEN-HEY group increased significantly.Transfected cells were inoculated into nude mice,and there were transplanted tumors at inoculated sites of all mice,and tumor growth in the PTEN-HEY group was significantly lower than that in the pcDNA-HEY group,and the weight was significantly lower than that in the pcDNA-HEY group.At 4th,5th and 6th day of the cells cultured with 4 μg/ml cisplatin,cell survival rate in the PTEN-HEY group was significantly lower than those in the HEY and pcDNA-HEY groups.Conclusions Up-regulating PTEN gene expression can promote apoptosis in HEY of human ovarian cell line,and increase the sensitivity to cisplatin chemotherapy.
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Objective To investigate the roles of phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway and its regulatory protein peroxisome proliferator-activated receptor γ (PPAR γ) and phosphatase and tension homologue deleted on chromosome 10 (PTEN) in regulating insulin sensitivity in rats with fetal growth restriction (FGR).Methods Sixteen pregnant rats were randomly divided into two groups including FGR and control groups on the 12th day of pregnancy (eight in each group).The FGR group was given low protein diet (8％ of casein) and restriction diet to establish the neonatal rat model of FGR.All maternal rats after delivery and newborn rats after weaning on 21 days after born were fed with normal diet.Each time blood samples were collected from eight newborn rats of each group to measure levels of fasting plasma glucose (FPG) and fasting insulin(FINS) at the time points of 21 days,two and four months after birth.Then insulin resistance index (IRI) and insulin sensitivity index (ISI) were calculated to evaluate insulin sensitivity.Expression of PI3K,AKT,PPAR γγ,PTEN and glucose transporters 4 (GLUT4) in skeletal muscle at mRNA and protein levels were measured at 21 days,two and four months after birth with real time fluorescence polymerase chain reaction and Western blot,respectively.Relationships between the expression of key molecules of PI3K/AKT signaling pathway and insulin sensitivity were analyzed.T-test,and Pearson's correlation analysis were used for statistical analysis.Results (1) The average birth weight of newborn rats in the FGR group was lower than that of the control group [(4.37± 0.69) vs (7.03±0.55) g,t=-20.75,P＜0.05].The incidence of FGR in the FGR group was 93.33％ (70/75).(2) Compared with normal offspring,those in the FGR group showed significantly increased FPG [two months after birth:(5.53± 0.58) vs (7.49 ± 0.38) mmol/L,t=8.08;four months afterbirth:(6.35±0.66) vs (8.94±0.90) mmol/L,t=6.58],FINS [two months afterbirth:(9.18±0.66) vs (14.67± 1.90) mU/L,t=7.71;four months after birth:(33.08±2.76) vs (56.33±2.81) mU/L,t=16.71] and IR1 (two months after birth:2.25±0.31 vs 4.90±0.81,t=8.63;four months after birth:9.30±0.90 vs 22.44±3.10,t=1 1.51),but decreased ISI (two months after birth:0.020 ± 0.002 vs 0.009± 0.001,t=-10.1 4;four months after birth:0.005±0.000 vs 0.002 ±0.000,t=-14.91) at two and four months after birth (all P＜0.05).(3) Compared with normal offspring,those in the FGR group showed decreased expression of PI3K (21 days after birth:0.082±0.028 vs 0.019±0.004,t=-6.29;two months after birth:0.020±0.003 vs 0.010±0.005,t=-4.78;four months after birth:0.014±0.004 vs 0.003±0.001,t=-7.87) and GLUT4 (21 days after birth:0.132±0.057 vs 0.041 ±0.019,t=-4.32;two months after birth:0.183±0.084 vs 0.069±0.017,t=-3.74;four months after birth:0.248±0.069 vs 0.113±0.040,t=-4.74) at mRNA level at 21 days,two and four months after birth (all P＜0.05).Compared with normal offspring,decreased expression of PPAR γ (two months after birth:0.028±0.002 vs 0.012±0.005,t=-3.70;four months after birth:0.030±0.008 vs 0.012±0.005,t=-3.80) and increased expression of PTEN (two months after birth:0.020±0.004 vs 0.045±0.014,t=5.09;four months after birth:0.023±0.007 vs 0.034±0.009,t=2.57) at mRNA level were observed in offspring of the FGR group at two and four months after birth (all P＜0.05).(4) Compared with normal offspring,expression of PI3K protein (21 days after birth:0.22±0.01 vs 0.17±0.02,t=-6.62;two months after birth:0.27±0.03 vs 0.16±0.02,t=-7.25;four months after birth:0.18±0.01 vs 0.09±0.02,t=-9.79) and GLUT4 protein (21 days after birth:0.21 ±0.01 vs 0.03±0.01,t=-27.29;two months after birth:0.10±0.01 vs 0.06t±0.01,t=-3.90;four months after birth:0.13 ±0.01 vs 0.08± 0.02,t=-8.10) decreased in offspring in the FGR group at 21 days,two and four months after birth (all P＜0.05).Compared with normal offspring,those in the FGR group showed decreased expression of PPAR γ protein (two months after birth:0.10 ± 0.01 vs 0.07± 0.01,t =-7.29;four months after birth:0.09±0.01 vs 0.08±0.01,t=-2.83),but increased expression of PTEN at protein level (two months after birth:0.10±0.01 vs 0.15±0.02,t=6.01;four months after birth:0.09±±0.01 vs 0.13±0.02,t=5.51) at two and four months after birth (all P＜0.05).(5) The IRI levels in offsprings in the FGR group were negatively correlated with the expression of PI3K,GLUT4 and PPAR γ at protein level (two months after birth:r=-0.90,-0.92 and-0.79;four months after birth:r=-0.92,-0.75 and-0.73,all P＜0.05),but positively correlated with the expression of PTEN at protein level (r=0.87 and 0.86,both P＜0.05) at two and four months after birth.Conclusions The abnormal expression of the key molecules of PI3K/AKT signaling pathway precedes the decrease of insulin sensitivity in newborn rats with FGR and the expression regulatory protein PPAR γ and PTEN are also changed,suggesting that these molecules may induce the impairment of insulin sensitivity in rats with FGR and be involved in the development of insulin resistance.
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Objective To investigate the effects of abated microRNA-21 (miRNA-21) on phosphatase and tensin homologue on chromosome ten protein (PTEN) and PI3K/Akt/mTOR pathway,as well as their further influence on the autophagy in high glucose (HG,25.0 mmol/L) induced rat glomerular mesangial cells.Methods MiRNA-21 inhibitor and negative control were transfected by liposome 2000 into rat glomerular mesangial cells (HBZY-1).The cells were divided into normal glucose (5.5 mmol/L) group,normal glucose + negative control group,normal glucose +miRNA-21 inhibitor group,HG group,HG+negative control group and HG+miRNA-21 inhibitor group.Cell proliferation and hypertrophy were assayed by MTT and the ratio of total protein to cell number respectively.The miRNA-21 expression was detected using real time PCR.The expressions of PTEN/ Akt/mTOR signaling signatures,autophagy-associated protein (p62 and LC3 Ⅱ) and collagen Ⅰ was detected by Western blotting and real time PCR.Autophagosomes were observed using electron microscopy.Results Compared with those in normal glucose group,in HG group cells had hypertrophy and proliferation,up-regulated miRNA-21 expression,and down-regulated PTEN protein and mRNA expressions (all P ＜ 0.01).Also there were and up-regulated p-Akt,p-mTOR,p62 and collagen Ⅰ expression,and lower LC3 Ⅱ expression and autophagosomes (all P ＜ 0.01).Further,compared with those in HG group,cells hypertrophy and proliferation in HG+miRNA-21 inhibitor group were reduced,expressions of p-Akt,p-mTOR,p62 and collagen Ⅰ were down-regulated,while expressions of PTEN and LC3 Ⅱ and autophagosomes were up-regulated (all P ＜ 0.01).Conclusions MiRNA-21 inhibitor up-regulates PTEN expression,which inhibits the activation of Akt/mTOR signaling pathway,ameliorates cell hypertrophy,proliferation and enhances autophagy to reduce extracellular matrix accumulation.
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Objective@#To investigate the effects of abated microRNA-21 (miRNA-21) on phosphatase and tensin homologue on chromosome ten protein (PTEN) and PI3K/Akt/mTOR pathway, as well as their further influence on the autophagy in high glucose (HG, 25.0 mmol/L) induced rat glomerular mesangial cells.@*Methods@#MiRNA-21 inhibitor and negative control were transfected by liposome 2000 into rat glomerular mesangial cells (HBZY-1). The cells were divided into normal glucose (5.5 mmol/L) group, normal glucose+negative control group, normal glucose+miRNA-21 inhibitor group, HG group, HG+negative control group and HG+miRNA-21 inhibitor group. Cell proliferation and hypertrophy were assayed by MTT and the ratio of total protein to cell number respectively. The miRNA-21 expression was detected using real time PCR. The expressions of PTEN/Akt/mTOR signaling signatures, autophagy-associated protein (p62 and LC3 Ⅱ) and collagen Ⅰ was detected by Western blotting and real time PCR. Autophagosomes were observed using electron microscopy.@*Results@#Compared with those in normal glucose group, in HG group cells had hypertrophy and proliferation, up-regulated miRNA-21 expression, and down-regulated PTEN protein and mRNA expressions (all P<0.01). Also there were and up-regulated p-Akt, p-mTOR, p62 and collagen Ⅰ expression, and lower LC3 Ⅱ expression and autophagosomes (all P<0.01). Further, compared with those in HG group, cells hypertrophy and proliferation in HG+miRNA-21 inhibitor group were reduced, expressions of p-Akt, p-mTOR, p62 and collagen Ⅰ were down-regulated, while expressions of PTEN and LC3 Ⅱ and autophagosomes were up-regulated (all P<0.01).@*Conclusions@#MiRNA-21 inhibitor up-regulates PTEN expression, which inhibits the activation of Akt/mTOR signaling pathway, ameliorates cell hypertrophy, proliferation and enhances autophagy to reduce extracellular matrix accumulation.
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Objective@#To study the expression status and clinical significance of PTEN and NDRG1 in colorectal carcinoma.@*Methods@#Tissue samples of 91 colorectal cancers, 30 colorectal adenomas and 21 colorectal normal mucosa tissues were collected. Postoperative specimens were examined by immunohistochemistry for PTEN and NDRG1 expression. The expression of PTEN and NDRG1 was correlated with clinicopathological feature.@*Results@#The expression of PTEN and NDRG1 in the studied cases was detected in 55.0%(50/91) and 76.9%(70/91), respectively. Their expression was significantly different from that of colorectal adenomas and normal colorectal mucosa tissues(P<0.05). Decreased expression of PTEN and over expression of NDRG1 were significantly related to the lymph node metastasis (P<0.05). The expression of PTEN was negatively related to that of NDRG1 in colorectal carcinoma(rs′=-0.251, P=0.016). The patients with negative expression of PTEN showed a lower disease free survival and overall survival(P<0.05).@*Conclusions@#Loss of expression of PTEN protein may be an important molecular marker in predicting the occurrence and PTEN may be useful as a prognostic marker of colorectal carcinoma. NDRG1 plays a role in the development of colorectal carcinoma, although not a prognostic indicator.The ancillary study with combined detection of PTEN and NDRG1 may be useful in difficult cases.
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Objective To explore the roles of miR-10a in the cisplatin resistance in cervical cancer cell lines,and further study the mechanism.Methods The cells were transfected with the mimics of miR10a and its negative control RNA (NC).Methyl thiazolyl tetrazolium (MTF) assay and fluorescence activated cell sorter were used to analyze drug sensitivity and apoptosis after treating with cisplatin,luciferase reporter assay to identify that miR-10a directly targets phosphatase and tensin homologue deleted on chromosome ten (PTEN).The expression of PTEN gene and its protein levels after tansfection was measured,respectively.Results after transfected with miR-10a mimics:(1) The 50％ inhibition concentration of Hela cells (7.2 μg/ml) was more than in the NC group (5.6 μg/ml).The 50％ inhibition concentration of Siha cells (6.4 μg/ml) was significantly more than in the NC group (3.8 μg/ml) (P＜0.05).(2) The apoptosis rates of Hela cells and Siha cells were significantly lower compared to the NC group (P ＜ 0.05).(3)miR-10a might directly target the 3'-untranslated region (3'-UTR) of PTEN,which significantly changed the PTEN protein level (P ＜ 0.05).Conclusions miR-10a may modulate cisplatin-induced apoptosis in cervical cancer by inhibiting PTEN protein level.
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Objective: To detect the methylation status of phosphatase and tensin homology deleted on chromosome ten (PTEN ) gene promoter in triple-negative breast cancer, and to analyze the correlation of PTEN gene methylation with the clinical and pathological characteristics of breast cancer patients. Methods: All of sixty cases of triple-negative breast cancer tissue specimens and sixteen cases of fresh normal breast tissue specimens (as the control) were selected. The methylation rate of PTEN gene promoter in the two groups was detected by pyrophosphate sequencing method. The expression levels of Ki-67, E-cadherin, vascular endothelial growth factor (VEGF), p53, and epidermal growth factor receptor (EGFR) were determined by immunohistochemical method. Then the relationships of PTEN gene methylation rate and the different clinical and pathological parameters of patients with triple-negative breast cancer were statistically analyzed. Results: The methylation rate of PTEN gene promoter in triple-negative breast cancer tissues was significantly higher than that in normal breast tissues (P < 0.05). For the patients with triple-negative breast cancer, the methylation rate of PTEN gene in cancer tissues at stage ? was higher than that at stage 0-? (P < 0.01), the one with lymph node metastasis was higher than that without lymph node metastasis (P < 0.01). Moreover, the methylation rates of PTEN gene in triple-negative breast cancer tissues with medium and high expressions of VEGF and p53 were higher than those with negative and low expressions of VEGF and p53 (both P < 0.01), and the one with Ki-67 level ≥ 14 % was higher than that with Ki-67 < 14 % (P < 0.01). Conclusion: The methylation status of PTEN gene promoter in triple-negative breast cancer tissues may be related with the clinical stage, lymph node metastasis and the expressions of VEGF, p53 and Ki-67 proteins, suggesting that PTEN gene methylation may play an important role in the occurrence and development of triple-negative breast cancer.
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ABSTRACT NKX3.1 and PTEN genes are involved in the development and progression of prostate cancer (PCa). Here, in line with other studies that correlated the expression of these two genes, we aimed at evaluating the expression pattern of these genes in clinical PCa samples. Collectively, 81 tissue samples including 45 human PCa and 36 benign prostatic hyperplasia (BPH) specimens were included in the study. The tissue samples were subjected to RNA extraction and subsequently to cDNA synthesis according to the kit manufacturer's protocol. Quantitative Real-Time PCR assay was performed for each sample in triplicate reactions. REST and SPSS software were used to statistically analyze PTEN and NKX3.1 gene expression data. Expression level of both NKX3.1 and PTEN genes was down-regulated in PCa samples compared to BPH samples. The relative expression ratio of PTEN and NKX3.1 was decreased to 0.155 and 0.003, respectively (P=0.000). The results of Chi-Square analysis revealed a significant correlation between the expression of these genes in both BPH and cancer groups (P=0.004 and 0.001, respectively). According to previous studies and our data, we concluded that the association between the down-regulation of PTEN and NKX3.1 genes contributed to the prostate tumorigenesis. This might highlight the interaction between the proteins encoded by these genes. Furthermore, this finding might be exploited for the development of innovative diagnostic and therapeutic approaches in PCa.