MiR-506通过PI3K/AKT信号通路调控人绒毛膜癌细胞增殖、迁移和侵袭

目的 探讨MiR-506是否通过PI3K/AKT通路对人绒毛膜癌细胞JEG-3的增殖、迁移、侵袭和凋亡发挥调控作用。方法 采用qRT-PCR法测定绒毛膜滋养层细胞HTR8/Svneo和绒毛膜癌细胞JEG-3中MiR-506的表达水平,将JEG-3细胞分组后转染MiR-506模拟物或抑制物后进行培养,采用CCK-8法测定细胞增殖情况,采用流式细胞术测定各组细胞凋亡情况,采用Western blot法测定PI3K/AKT通路相关蛋白表达情况。结果JEG-3细胞中MiR-506表达水平较HTR8/Svneo细胞降低;MiR-506模拟物组JEG-3细胞中MiR-506表达水平升高,MiR-506抑制物组MiR-506表达水平降低;MiR-506模拟物组24 h、48 h、72 h时吸光度增加,而MiR-506抑制物组吸光度显著降低;MiR-506模拟物组细胞凋亡数量升高（P <0.05）,而MiR-506抑制物组细胞凋亡数量降低（P <0.05）;MiR-506模拟物组PI3K和AKT蛋白表达水平降低（P <0.05）,MiR-506抑制物组PI3K和AKT蛋白表达水平增加...     

MiR-506通过PI3K/AKT信号通路调控人绒毛膜癌细胞增殖、迁移和侵袭#br#

Objective?To investigate the regulation of miR-506 on proliferation, migration, invasion and apoptosis of choriocarcinoma JEG-3 cells via PI3K/AKT signal pathway. Methods?qRT-PCR was used to determine miR-506 in JEG-3 cells and normal HTR8/Svneo cells. JEG-3 cells were cultured after transfection and divided into different groups. CCK 8 method was used to determine cell proliferation. Flow cytometry was used to determine cell apoptosis. Western-blot were used to determine and protein related to PI3K/AKT signal pathway. Results?The expression of miR-506 in JEG-3 cells was increased compared with that in HTR8/Svneo cells; miR-506 in miR-506 mimics group increased, while MiR-506 in miR-506 inhibitor group decreased. The absorbance value in miR-506 mimic group at 24 h, 48 h and 72 h decreased , while absorbance value in miR-506 inhibitor group at 24 h, 48 h and 72 h increased. The apoptosis rate in miR-506 group increased, while the apoptosis rate in miR-506 inhibitor group decreased. The mRNA and protein levels of PI3K and AKT in miR-506 mimic group decreased, while the mRNA and protein levels of PI3K and AKT in miR-506 inhibitor group increased. Conclusion?MiR-506 is a tumor suppressor gene of choriocarcinoma, which may regulate the proliferation, migration, invasion and apoptosis of choriocarcinoma of choriocarcinoma cells via the PI3K/AKT signal pathway.     

芬太尼通过PI3 K/Akt信号通路调控宫颈癌细胞增殖迁移

目的:探究芬太尼对宫颈癌细胞增殖迁移的作用及分子机制.方法:将宫颈癌细胞系C33 A分为对照组和芬太尼组(使用不同浓度芬太尼处理),分别采用CCK-8、Transwell法、流式细胞术检测芬太尼对C33 A细胞增殖、迁移及凋亡的影响.Western blot法检测活化Caspase 3、PARP1含量,MMP-2、MM...     

PI3K/Akt/mTOR信号通路靶向治疗在子宫内膜癌中的研究进展

子宫内膜癌是女性高发的恶性肿瘤之一,在生殖系统中常见的恶性肿瘤位居第三,近年其发病率呈上升趋势。磷磷脂酰肌醇-3-激酶/蛋白激酶B/雷帕霉素靶蛋白(PI3K/Akt/m TOR)信号通路是子宫内膜癌中发生最频繁的信号转导通路之一。有研究证实,24%~39%的子宫内膜癌患者存在PI3K/Akt/m TOR信号通路的异常激活。众多研究表明,其通路中分子的改变不同程度的参与并影响了多种生物功能,PI3K/Akt/m TOR信号通路的抑制剂在子宫内膜癌治疗中也起到了重要作用,并引发了其机制研究的热潮。临床前期实体肿瘤和临床各期试验研究均在证明不同种类PI3K/Akt/m TOR信号通路抑制剂的作用。本文将对子宫内膜癌中PI3K/Akt/m TOR信号通路整体阐述,进而对该通路各个组件相关的抑制剂做总结,从而对于药物靶向治疗或联合其他方案治疗子宫内膜癌提供广阔的治疗思路。     

PI3K/PKB信号传导通路与子宫内膜癌研究进展

磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(PKB)信号通路是许多生长因子、细胞因子等的下游信号传导通路,通过抑制细胞凋亡,促进细胞生长、增殖、迁移、侵袭和肿瘤血管生成而与肿瘤发生发展密切相关。许多与子宫内膜癌相关的因素通过PI3K/PKB通路参与肿瘤发生发展,这为子宫内膜癌病因学研究及寻找新的治疗靶点提供了新思路。     

PI3K/PKB信号传导通路与子宫内膜癌研究进展

磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(PKB)信号通路是许多生长因子、细胞因子等的下游信号传导通路,通过抑制细胞凋亡,促进细胞生长、增殖、迁移、侵袭和肿瘤血管生成而与肿瘤发生发展密切相关.许多与子宫内膜癌相关的因素通过PI3K/PKB通路参与肿瘤发生发展,这为子宫内膜癌病因学研究及寻找新的治疗靶点提供了新思路.     

双酚A激活PI3K/AKT通路促进子宫内膜癌细胞增殖的研究

目的:研究双酚A(bisphenol A,BPA)活化磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)促子宫内膜癌细胞Ishikawa和RL952增殖的机制。方法:CCK8试剂盒检测Ishikawa和RL952细胞的增殖情况,蛋白质印迹(Western blotting)法检测p-AKT蛋白的表达。结果:在Ishikawa和RL952细胞中,BPA作用48 h时,细胞增殖呈浓度依赖性,1μmol/L的BPA促细胞生长效应最显著,Ishikawa和RL952细胞增殖率分别为0.758±0.023和0.692±0.042。BPA浓度超过1μmol/L后,促细胞增殖的作用下降。BPA作用24 h时促增殖效应不明显,BPA作用72 h时表现出细胞毒作用。1μmol/L BPA处理的Ishikawa和RL952细胞48 h后p-AKT蛋白的表达较对照组升高(P<0.05),加入PI3K抑制剂(LY294002),p-AKT的蛋白表达比对照组降低,但差异无统计学意义(P>0.05)。结论:BPA通过激活PI3K/AKT通路促进子宫内膜癌细胞增殖。     

PI3K/Akt/mTOR和MAPK信号通路与子宫内膜异位症的研究进展

子宫内膜异位症(EMs)是常见的妇科良性疾病,却具有侵袭、转移等特性。PI3K/Akt/m TOR和MAPK信号通路是两条重要的细胞内信号转导通路,在多种细胞反应中发挥着重要作用。研究显示,在EMs中存在PI3K/Akt/m TOR和MAPK信号通路的过度激活,并通过调节细胞黏附分子(CAM)、基质金属蛋白酶(MMP)、血管内皮生长因子(VEGF)及凋亡因子等的表达,影响异位内膜的黏附、侵袭,血管生成及凋亡,并受激素水平的调控,促进了子宫内膜细胞的异位侵袭和转移,与EMs的发生、发展密切相关。     

抑制PI3K/Akt信号通路拮抗胰岛素诱导的子宫内膜癌细胞增殖

目的:研究抑制磷脂酰肌醇3激酶(PI3K)/Akt信号通路对胰岛素诱导的子宫内膜癌细胞增殖的拮抗作用。方法:将无血清饥饿的子宫内膜癌Ishikawa3-H-12细胞分为空白对照组、10-6mol/L胰岛素单独刺激组以及不同剂量PI3K抑制剂-LY294002预处理后再用胰岛素刺激组。Western blot检测各组Akt磷酸化(p-Akt)水平,MTT试验观察细胞增殖情况。结果:胰岛素可引起内膜癌细胞Akt活化,刺激15min后p-Akt/Akt比值显著高于空白对照组(68.68%vs 26.21%,P<0.001)。LY294002以浓度依赖方式抑制胰岛素引起的Akt磷酸化。MTT试验显示,在药物处理24h,48h和72h 3个时间点,不同组别570nm吸光度值(OD570nm)均有显著差异(F=156.329,700.973,812.224,均P<0.001)。胰岛素组OD570nm值均高于同时间点的空白对照组(均P<0.001),胰岛素促内膜癌细胞增殖作用于48h时最为显著。LY294002可抑制胰岛素的增殖促进作用,此抑制作用具有浓度依赖性。不同剂量LY294002抑制作用的时间依赖性不同,48h时小剂量(0.1、1、10μmol/L)的抑制作用最为显著,72h时胰岛素重新呈现一定的促增殖作用;而50μmol/L LY294002可以持久抑制胰岛素的促增殖作用。结论:PI3K抑制剂LY294002可以通过抑制Akt磷酸化阻断胰岛素信号传导,拮抗后者促子宫内膜癌细胞增殖的作用。     

PI3K/Akt信号通路与卵巢早衰相关性的研究进展

近年研究发现,PI3K/Akt信号通路参与卵巢早衰(POF)疾病发生。磷脂酰肌醇3-激酶(PI3K)和蛋白激酶B(Akt)的过度激活可使原始卵泡过早发育及卵泡过快凋亡,进而发展成为POF。本文就关于PI3K/Akt信号通路与POF关系的研究进展做一综述。     

Metformin is associated with reduced cell proliferation in human endometrial cancer by inbibiting PI3K/AKT/mTOR signaling

Metformin recently gained traction as potential anti-endometrial cancer agent for its new applications. However, the underlying mechanisms of the anti-cancer effect of metformin in the endometrial cancer have not yet been fully elucidated. Sixty-five patients diagnosed as endometrial carcinoma were grouped into (n?=?33) and non-treatment mixed (n?=?32) for analysis. Thirty healthy donors were recruited as controls. We attempt to investigate the effect of metformin on Ki-67, PI3K, p-AKT, p-S6K1, and p-4EBP1 staining in human endometrial cancer by immunohistochemical staining. We found that increased Ki-67 expression in women with endometrial cancer, which were reversed by conventional anti-diabetic doses of metformin in present work. In parallel, the reduced PI3K, p-AKT, p-S6K1, and p-4EBP1 staining induced by metformin appeared to play an important role for the anti-proliferative effects of metformin in endometrial cancer patients. Metformin significantly decreased proliferation in human endometrial cancer may by inhibiting PI3K/AKT/mTOR signaling. Our present results add to the growing body of evidence supporting metformin as a potential anti-cancer agent in endometrial cancer.     

Assessing the efficacy of targeting the phosphatidylinositol 3-kinase/AKT/mTOR signaling pathway in endometrial cancer

Objective

Alterations in the PI3K pathway are prevalent in endometrial cancer due to PIK3CA mutation and loss of PTEN. We investigated the anti-tumor activity of the PI3K inhibitor NVP BKM-120 (BKM) as a single agent and in combination with standard cytotoxic chemotherapy in a human primary endometrial xenograft model.

Methods

NOD/SCID mice bearing xenografts of primary human tumors with and without PIK3CA gene mutations were divided into two and four arm cohorts with equivalent tumor volumes. BKM was administered alone and in combination with paclitaxel and carboplatin (P/C) and endometrial xenograft tumor volumes were assessed. Tumors from the BKM, P/C, P/C + BKM and vehicle treated mice were processed for determination of PI3K/AKT/mTOR pathway activation.

Results

In both single agent experiments, BKM resulted in significant tumor growth suppression starting at days 5–10 compared to the linear growth observed in vehicle treated tumors (p < 0.04 in all experiments). Tumor resurgence manifested between days 14 and 25 (p < 0.03). When BKM was combined with P/C, this resistance pattern failed to develop in three separate xenograft lines (p < 0.05). Synergistic tumor growth suppression (p < 0.05) of only one xenograft tumor with no detected PIK3CA mutation was observed. Acute treatment with BKM led to a decrease in pAKT levels.

Conclusion

Independent of PIK3CA gene mutation, BKM mediated inhibition of the PI3K/AKT/mTOR pathway in endometrial tumors precludes tumor growth in a primary xenograft model. While a pattern of resistance emerges, this effect appears to be mitigated by the addition of conventional cytotoxic chemotherapy.     

Normal endometrial stromal cells regulate survival and apoptosis signaling through PI3K/AKt/Survivin pathway in endometrial adenocarcinoma cells in vitro

Objective

Stroma–tumor communication plays an important role in the genesis of neoplasia. In the current study, we investigated the effect of normal stromal cells on the survival and apoptosis signaling of endometrial cancer cells and further explored the possible mechanism implied in this communication.

Methods

Using primarily cultured normal endometrial stromal cells and an endometrial adenocarcinoma cell line, Ishikawa cells, we established a 2D-coculture system to observe the stromal cell–tumor cell crosstalk in endometrial carcinomas. Using methyl thiazolyl tetrazolium (MTT) assays, cell counting and colony formation assays, we analyzed the effect of stomal cells on the growth and proliferation of Ishikawa cells under different conditions. Using western blot analysis, we determined the effect of stromal cells on the activity of PI3K/AKt/Survivin signaling in Ishikawa cells under different conditions. Using immunohistochemistry analysis, we determined the expression of Survivin in normal endometria and endometrial adenocarcinomas.

Results

We found that the paracrine factors from normal endometrial stromal cells grown on Matrigel repeatedly and significantly decreased hormone-stimulated activity of PI3K/AKt/Survivin signaling in Ishikawa cells, which were proved to be increased in endometrial adenocarcinoma and essential in hormone-induced cell growth in Ishikawa cells.

Conclusion

Paracrine factors from normal endometrial stromal cells can inhibit hormone-stimulated cell proliferation in Ishikawa cells by regulating cell survival and apoptosis through PI3K/AKt/Survivin signaling.     

沉默PI3K基因对E2刺激后子宫内膜癌细胞增殖和凋亡的影响

目的:探讨沉默PI3K基因对雌激素刺激下的子宫内膜癌细胞增殖、迁移、凋亡的影响。方法:培养子宫内膜癌Ishikawa细胞,构建LV-PI3K-RNAi慢病毒载体转染Ishikawa细胞。实验分组Control组未经任何处理的Ishikawa细胞;Ishikawa组经E₂刺激的Ishikawa细胞;NC组经E₂刺激并转染阴性对照慢病毒的Ishikawa细胞;PI3Ki组经E₂刺激并转染LV-PI3K-RNAi慢病毒的Ishikawa细胞。Real-time PCR、Western blot法检测各组VEGF、bFGF、PI3K mRNA及蛋白表达水平,MTT法、流式细胞仪分别检测细胞增殖能力及凋亡的情况。结果:与Ishikawa组、NC组比较,PI3Ki组的VEGF、bFGF、PI3K mRNA及蛋白表达水平明显降低,细胞增殖能力显著降低,凋亡率增高,差异均有统计学意义(P<0.05)。结论:PI3K基因低表达可干扰E₂在基因、蛋白水平激活Ishikawa细胞产生VEGF、bFGF,从而下调E₂对子宫内膜癌细胞增殖和抑制凋亡的影响。     

SHBG expression is correlated with PI3K/AKT pathway activity in a cellular model of human insulin resistance

Decreased sex hormone-binding globulin (SHBG) expression is an independent risk factor for gestational diabetes mellitus(GDM).However, the mechanisms that link low SHBG expression and insulin resistance in GDM is unclear. In this study, we investigated the placenta SHBG in the PI3K/AKT pathway to reveal the mechanism that links decreased SHBG to insulin resistance. A insulin resistance cells model was established by the method of insulin stimulation. Two groups were set up, HTR8/Svneo cells and insulin-resistance cells of HTR8/SVneo. The expression of SHBG and PI3K/AKT associated factors were detected using real-time PCR and western blotting and their correlations were analyzed. The results showed that SHBG protein and mRNA levels in insulin resistance cells were both significantly lower. Along with decreased SHBG expression, the mRNA and protein levels of IRS-1, IRS-2, PI3Kp85α and GLUT-3, GLUT-4 decreased significantly. However, the expression of GLUT-1 increased significantly. Pearson correlation analysis showed that SHBG mRNA expression was positively correlated with IRS-1, IRS-2 and PI3Kp85α mRNA levels. According to the results, low SHBG expression not only participates in the development of local insulin resistance, but may also play an important role in PI3K/AKT pathway-mediated systemic insulin resistance and gestational diabetes.     

The role of the phosphatidylinositol 3-kinase (PI3K) pathway in the development and treatment of uterine cancer

Objective

Uterine cancer is the most common gynecologic malignancy in the United States. Although surgery is often curative for women diagnosed in the early stages, prognosis for patients with advanced disease is poor. Alterations in the phosphatidylinositol 3-kinase (PI3K) pathway are known to play a significant role in the development of uterine cancer and provide a possible target for new therapies.

Methods

PubMed was searched for articles of relevance to uterine cancer and the PI3K pathway. In addition, abstracts from key oncology congresses were scanned for data on novel therapeutic agents targeting the PI3K pathway.

Results

The PI3K pathway is an important promoter of cellular growth, metabolism, differentiation, proliferation, survival, and angiogenesis. It is often upregulated in uterine cancer, with the most frequent genetic alterations occurring in phosphatase and tensin homolog (PTEN), PIK3CA, and KRAS. Deregulation of the pathway has been associated with resistance to hormonal therapy and chemotherapy. Inhibitors of the PI3K pathway are in clinical testing in patients with solid tumors, including uterine cancer. Results with monotherapy demonstrate some clinical responses, but mainly as prolonged stable disease. PI3K pathway inhibitors are currently being evaluated in patients with tumors in which the PI3K pathway is deregulated. Another strategy being evaluated is the ability of PI3K pathway inhibitors to restore sensitivity to standard therapy.

Conclusions

Investigational PI3K pathway inhibitors represent a promising new therapeutic strategy in uterine cancer. Exploration of effective drug combinations and their applicability to individual tumors will be important in the future clinical development of these agents.     

Effect of peritoneal fluid from patients with mild and severe endometriosis on endometrial stromal cell proliferation

The aim of this study was to evaluate and compare the mitogenic effect of peritoneal fluid (PF) from women with mild and severe endometriosis on the endometrial stromal cell proliferation. Increasing concentrations of PF from women with and without mild or severe endometriosis were added to primary endometrial stromal cell cultures and³H-thymidine incorporation was used to assess DNA synthesis in these cultures. PF from women with mild endometriosis induced a statistically significant dose-dependent increase in stromal cell thymidine uptake ranged from 5.8 to 14.5 fold, whereas PF from women with severe endometriosis produced an average 51% inhibition of stromal cell proliferation of compared with cells exposed to non-endometriosis PF or exposed to nutrient medium supplemented with 2.5% calf serum alone. PF samples from patients with stage I endometriosis induced a statistically dose-dependent increase in stromal cell proliferation, whereas PF from patients with stage IV endometriosis caused a significant inhibition.