PTEN基因和P53基因在子宫内膜癌中的表达及意义

目的：探讨PTEN基因和P53基因在子宫内膜癌中的表达及临床意义,为子宫内膜癌的治疗提供参考依据。方法：用免疫组化法检测50例子宫内膜癌患者PTEN基因缺失和P53基因突变情况,并分析两者与子宫内膜癌不同病理变化的相关性。结果：PTEN缺失24例,缺失率48.00%,进一步研究显示PTEN缺失与细胞分化程度和肌层浸润情况密切相关（P〈0.05）,P53突变32例,缺失率64.00%,并进一步研究显示P53突变与FIGO分期,细胞分化程度和肌层浸润情况密切相关（P〈0.05）。结论：PTEN基因缺失和P53基因突变与子宫内膜癌发生和发展密切相关,以PTEN基因和P53基因为靶点的生物治疗在进展期内膜癌中的治疗价值值得进一步深入研究。     

PTEN基因和P53基因在子宫内膜癌中的表达及意义

目的:探讨PTEN基因和P53基因在子宫内膜癌中的表达及临床意义,为子宫内膜癌的治疗提供参考依据。方法:用免疫组化法检测50例子宫内膜癌患者PTEN基因缺失和P53基因突变情况,并分析两者与子宫内膜癌不同病理变化的相关性。结果:PTEN缺失24例,缺失率48.00%,进一步研究显示PTEN缺失与细胞分化程度和肌层浸润情况密切相关(P0.05),P53突变32例,缺失率64.00%,并进一步研究显示P53突变与FIGO分期,细胞分化程度和肌层浸润情况密切相关(P0.05)。结论:PTEN基因缺失和P53基因突变与子宫内膜癌发生和发展密切相关,以PTEN基因和P53基因为靶点的生物治疗在进展期内膜癌中的治疗价值值得进一步深入研究。     

子宫内膜癌中PTEN与PI3K-Akt通路的表达及意义

目的：研究PTEN与P1K3-Akt通路在子宫内膜癌中的表达及临床意义。方法：采用免疫组化技术检测正常子宫内膜组织（24例）、子宫内膜样腺癌组织（55例）及子宫内膜浆液性癌组织（26例）中PTEN、P13Kp110α及P—AKTSer473的表达情况,分析其表达与组织病理参数的关系。结果：免疫组化结果显示：子宫内膜癌的PTEN表达缺失率,P13Kp110α及p-AKTser473表达阳性率均高于正常子宫内膜组织,且正常子宫内膜组织与子宫内膜样腺癌组织比较有统计学意义（P〈0．05）;随着肌层浸润深度增加,PTEN表达缺失率增加（P〈0．05）。结论：PTEN的表达缺失及P13Kp110α与P-AKTSer473的表达可能与子宫内膜癌的发生发展有关,尤其是在子宫内膜样腺癌的发生发展机制中占重要地位。     

子宫内膜癌组织中PTEN和MTA1蛋白的表达及其相关性

目的：探讨子宫内膜癌组织中PTEN和MTA1表达及其与子宫内膜癌生物学行为之间的关系。方法：采用免疫组化SP法检测130例子宫内膜癌和40例正常宫内膜组织中PTEN和MTAl1的表达水平,并分析两者在子宫内膜癌中的相关性。结果：子宫内膜癌组织中PTEN、MTA1阳性表达均显著高于正常子宫内膜组织（P〈0．01）;PTEN与MTAI在子宫内膜癌组织中的表达呈负相关（r=0．35,P〈0．05）。结论：PTEN和MTA1表达与子宫内膜癌的发生、发展及生物行为密切相关,且两者表达存在负相关性。     

MiR-1297下调PTEN的表达参与子宫内膜癌变

已有研究证明,抑癌子PTEN（phosphatase and tensin homolog）缺失或表达下调可引起子宫内膜癌变;在喉鳞状细胞癌中miR-1297可下调PTEN的表达。然而,miR-1297下调PTEN的机制,以及是否miR-1297靶向抑制PTEN参与子宫内膜癌变尚未见报道。本研究证明,miR-1297靶向抑制PTEN基因的表达参与子宫内膜癌变。收集临床病理检查确诊为子宫内膜腺癌的癌组织及癌旁组织18例。实时定量PCR（RT-qPCR）及Western印迹结果显示,与癌旁子宫内膜比较,子宫内膜癌组织miR-1297高表达,而PTEN则呈低表达或缺失。内膜癌和癌旁内膜组织中,miR-1297与PTEN表达水平的散点图分析揭示,miR-1297的表达水平与PTEN的表达水平呈明显的负相关（内膜癌：miR-1297与PTEN的相关系数r=-0.6928,P=0.0438;癌旁组织：相关系数r=-0.4085,P < 0.0001)。报告酶活性测定显示,与对照比较,pGL3-PTEN-3′-UTR-1/2/3加miR-1297模拟物共转染后,荧光素酶活性在转染的Ishikawa和ECC-1细胞中分别降低了约21%（P=0.0196）和17%（P=0.0306）;相反,转染miR-1297抑制物后,两种细胞的荧光素酶活性分别升高约59%（Ishikawa：P=0.0014）和50%（ECC-1： P=0.0025）。结果提示,miR-1297可通过特异识别PTEN mRNA的3′-UTR,抑制PTEN的表达。基因转染结合Western印迹结果证明,在原代子宫内膜细胞中,过表达miR-1297模拟物导致PTEN蛋白表达下调,而其下游底物分子--磷酸化的AKT（p-AKT）表达上调;而过表达miR-1297抑制物后,PTEN表达上调,p-AKT表达下调。Connexin V/PI双染结合流式分析证明,与对照细胞（3%细胞凋亡）比较,在子宫内膜癌细胞系Ishikawa中,敲减miR-1297表达后细胞调亡明显增加（至10%）,这可能与抑制miR-1297后PTEN的表达上调有关。上述结果提示,miR-1297在子宫内膜癌高表达,而PTEN低表达。上述结果还提示,miR-1297作为一个促癌子可通过特异识别PTEN mRNA的3′-UTR,抑制PTEN的表达,促进AKT磷酸化/激活,从而促进内膜癌的发生,这可能是子宫内膜癌变的发生原因之一。     

PTEN蛋白在子宫内膜癌中的表达及其临床意义

目的　探讨抑癌基因PTEN产物PTEN蛋白在由正常子宫内膜过渡至子宫内膜癌过程中的表达缺失情况及其临床意义。方法　用SP免疫组化法测定 73例子宫内膜癌、 2 5例子宫内膜非典型增生、 71例子宫内膜增殖症和 31例正常子宫内膜组织中PTEN蛋白的表达 ,并与组织学类型、组织学分级、肌层浸润深度和临床分期等生物学行为进行相关分析。结果　PTEN蛋白在子宫内膜腺癌和癌前病变中的表达缺失率分别为 6 6 6 7%和 76 0 0 % ,表达缺失率与组织学类型(P <0 0 0 5 )和组织学分级有关 (P <0 0 5 ) ,与子宫内膜癌肌层浸润深度和临床分期无关 (P >0 0 5 )。结论　PTEN表达缺失是子宫内膜癌发生的早期分子事件 ,PTEN蛋白表达缺失是早期诊断子宫内膜癌及癌前病变较好的生物标志。     

子宫内膜癌组织中PTEN 和MTA1 蛋白的表达及其相关性

目的:探讨子宫内膜癌组织中PTEN和MTA1表达及其与子宫内膜癌生物学行为之间的关系。方法:采用免疫组化SP法检测130例子宫内膜癌和40例正常宫内膜组织中PTEN和MTA1的表达水平,并分析两者在子宫内膜癌中的相关性。结果:子宫内膜癌组织中PTEN、MTA1阳性表达均显著高于正常子宫内膜组织(P<0.01);PTEN与MTA1在子宫内膜癌组织中的表达呈负相关(r=-0.35,P<0.05)。结论:PTEN和MTA1表达与子宫内膜癌的发生、发展及生物行为密切相关,且两者表达存在负相关性。     

揭示抑癌基因抗病毒机制

正武汉大学生命科学学院与基础医学院郭德银研究组揭示了人类重要抑癌蛋白PTEN不仅具有抗肿瘤功能,还在抗病毒先天免疫中发挥关键作用,相关研究日前在线发表于《自然-免疫学》。Pten是一种重要的抗肿瘤基因,其突变或功能缺陷与多种肿瘤的发生直接相关,包括乳腺癌、子宫内膜癌、肝癌等,但其在机体抗病毒免疫方面是否能够发挥作用尚不清楚。此次研究发现,PTEN蛋白在抗病毒先天免疫中具有关键性作用,其通过调控决定Ⅰ型     

p53和VEGF蛋白表达与子宫内膜癌的关系

目的：检测子宫内膜癌组织中血管内皮生长因子与突变型抑癌基因p53的表达情况,以探讨其在子宫内膜癌发生转移中的作用。方法：应用免疫组化S．P法检测45例子宫内膜癌及10例正常子宫内膜组织中VEGF和突变型p53的表达及其相关性,分析其与各临床病理参数之间的关系。结果：子宫内膜癌组织中VEGF及突变型p53蛋白表达阳性率均高于正常子宫内膜,两者呈显著正相关（P〈0．01）。两者阳性表达率均与临床分期、组织分化程度、淋巴结转移（P〈0．05）有关,VEGF阳性表达率与肌层浸润深度无明显相关性（P〉0．05）,而突变型p53表达则与肌层浸润深度有关（P〈0.05）。结论：VEGF和突变型p53与子宫内膜癌的发生、侵袭、转移和预后相关。     

子宫内膜癌中TGF-β1及受体TβRI的表达及其临床意义

目的：研究转化生长因子（TGF-β1）及其Ⅰ型受体（TβRI）在子宫内膜癌组织中的表达及其临床意义。方法：采用RT-PCR方法检测36例子宫内膜癌及31例正常子宫内膜组织中TGF-β1及TβRI基因的表达水平,分析其与临床病理参数之间的关系。结果：子宫内膜癌组织中TGF-β1mRNA的表达明显高于正常组织（P〈0.05）,而TβRI mRNA的表达水平明显低于正常内膜组织（P〈0.05）。TGF-β1mRNA及TβRImRNA在子宫内膜癌中的表达均与肿瘤的临床分期、组织学分级、肌层浸润深度、淋巴结转移呈显著性相关（P〈0.05）,差别有统计学意义。结论：TGF-β1及其Ⅰ型受体TβRI在子宫内膜癌的发生、发展中可能起重要作用,检测TGF-β1及TβRI对评价子宫内膜癌的转移及预后有重要价值。     

Long noncoding RNA PTENP1 affects the recovery of spinal cord injury by regulating the expression of miR-19b and miR-21

Exosomes derived from differentiated P12 cells and MSCs were proved to suppress apoptosis of neuron cells, and phosphatase and tensin homolog pseudogene 1 (PTENP1) was reported to inhibit cell proliferation. In this study, we aimed to investigate the role of PTENP1 in the process of post-spinal cord injury (SCI) recovery, so as to evaluate the therapeutic effects of exosomes derived from MSCs transfected with PTENP1 short hairpin RNA (shRNA), as a type of novel biomarkers in the treatment of SCI. Electron microscopy was used to observe the morphology of different exosomes. Real-time polymerase chain reaction and western blot, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, flow cytometry, Nissl staining, immunohistochemistry assay, and terminal deoxynucleotidyl transferase dUTP nick end labeling assay were conducted to investigate and validate the underlying molecular signaling pathway. PTENP1-shRNA downregulated PTENP1 and PTEN while upregulating miR-21 and miR-19b. PTENP1-shRNA also accelerated cell apoptosis and reduced cell viability. In addition, PTENP1 reduced the miR-21 and miR-19b expression by directly targeting miR-21 and miR-19b. Meanwhile, both miR-21 and miR-19b reduced the expression of PTEN by directly targeting the 3′-untranslated region of PTEN. Furthermore, PTEN level and apoptosis index of neuron cells was the highest in the SCI group, while the treatment with exosomes+PTENP1-shRNA reduced the PTEN expression to a level similar to that in the sham group. Finally, PTENP1 inhibited miR-21 and miR-19b expression but upregulated PTEN expression. The upregulation of miR-21/miR-19b also suppressed the apoptosis of neuron cells by downregulating the PTEN expression. PTENP1 is involved in the recovery of SCI by regulating the expression of miR-19b and miR-21, and exosomes from PTENP1-shRNA-transfected cells may be used as a novel biomarker in SCI treatment.     

Isothiocyanate–drug interactions in the human adenocarcinoma cell line Caco-2

Loss or mutation of the PTEN (phosphatase and tensin homologue deleted on chromosome 10) gene is associated with resistance to epidermal growth factor receptor (EGFR) inhibitors. However, the mechanism underlying remains elusive. In this study, we aimed to explore whether sensitivity to the EGFR tyrosine kinase inhibitor (TKI) is affected by PTEN status in endometrial cancer cells. PTEN siRNA and the PTEN gene were transfected into HEC-1A and Ishikawa endometrial cancer cells using lentiviral vectors. Cells were treated under various concentrations of RG14620 and rapamycin, which are EGFR and mammalian target of rapamycin (mTOR) inhibitors, respectively. The IC(50) of RG16420 was determined by using the MTT method. Cell apoptosis and the cell cycle were studied, and activation of EGFR, AKT, and p70S6 were detected by Western blot analysis. Loss of PTEN promoted cell proliferation and led to significant increases in the levels of EGFR, phospho-EGFR, AKT, phospho-AKT, and phospho-mTOR proteins. Ishikawa and HEC-1A(PTENkd) cells that displayed loss and inactivation of PTEN function were resistant to RG14620. HEC-1A and Ishikawa(PTEN) cells with intact PTEN were sensitive to RG14620. The combination of two inhibitors was more effective than both monotherapies, particularly in carcinoma cells with PTEN dysfunction. Decreased phospho-EGFR protein expression was observed in all cell lines that were sensitive to RG14620. Decreased phospho-AKT and phospho-p70S6 protein expression was observed in PTEN-intact cells that were sensitive to RG14620. PTEN loss results in resistance to EGFR TKI, which was reversed by PTEN reintroduction or mTOR inhibitor treatment. The combined treatment of EGFR TKI and the mTOR inhibitor provided a synergistic effect by promoting cell death in PTEN-deficient and PTEN-intact endometrial cancer cells, particularly in PTEN-deficient carcinoma cells with up-regulated EGFR activation.     

Promoters of genes MTHFR from patients with hyperhomocysteinemia and PTEN from patients with malignant and benign endometrial and ovarian tumors

Mutational changes in the promoter regions of MTHFR genes from patients with hyperhomocysteinemia and PTEN genes from patients with endometrial and ovarian tumors were studied. An increased level of homocysteine was found in a part of the patients with a heterozygous C677T mutation in the MTHFR gene, although a moderate hyperhomocysteinemia is usually associated with homozygous mutation. We hypothesized that, in this case, the allele lacking the C677T mutation may be inactivated by the promoter mutation. The sequencing of both DNA strands of the minimal promoter region of the MTHFR gene in ten patients did not reveal any mutation, which implied another mechanism of the development of hyperhomocysteinemia in these patients. A PCR analysis of the minimal promoter region of the tumor suppressor PTEN in the presence of 2-pyrrolidone in 101 patients from Moscow clinics revealed changes in it in patients with endometrial (56%) or ovarian (29%) cancer, as well as in patients with endometrial hyperplasia and benign ovarian tumors (34.6 and 29%, respectively). It was presumed that the found PTEN gene promoters may arise from epigenetic alterations (erroneous methylation) or may (more rarely) be induced by mutations. As a result of the studies, new molecular markers associated with endometrial and ovarian tumors were revealed and a simple and effective method of detection of these markers was developed.     

Pseudogene PTENP1 sponges miR-214 to regulate the expression of PTEN to modulate osteoclast differentiation and attenuate osteoporosis

Background aimsOsteoporosis (OP) is a common bone metabolic disease with a high incidence. Our study aimed to explore the pseudogene PTENP1/miR-214/PTEN axis to modulate the osteoclast differentiation in osteoporosis.MethodsPatients with osteoporosis were recruited in our study, and RANKL-induced osteoclast differentiation and ovariectomy-induced osteoporosis mouse model were established in vitro and in vivo, respectively.ResultsPseudogene PTENP1 and PTEN were significantly down-regulated and miR-214 was up-regulated in osteoporosis patients. In addition, overexpression of PTENP1 or silence of miR-214 inhibited the expression levels of osteoclast specific markers and osteoclast differentiation induced by RANKL. Overexpression of PTENP1 or silence of miR-214 also inhibited the levels of phosphorylation of PI3K and AKT, p65 nuclear translocation, IκBα degradation and the expression level of NFATc1. AlsoSilence of PTENP1 or overexpression of miR-214 induced the osteoclast differentiation under normal physiological condition. Pseudogene PTENP1 sponged miR-214 to regulate the expression of PTEN.ConclusionsIn an ovariectomy-induced osteoporosis mouse model, obvious pathological changes in bone tissues were found, and bone marrow mononuclear cells in this group were more likely to differentiate into osteoclasts. Therefore, pseudogene PTENP1 sponged miR-214 to regulate the expression of PTEN to inhibit osteoclast differentiation and attenuate osteoporosis by suppressing the PI3K/AKT/NF-κB signaling pathway.