二甲双胍对人子宫内膜癌细胞迁徙能力的影响

目的：体外检测二甲双胍对人子宫内膜癌（Ec）细胞迁徙能力的影响,并探讨其可能的作用机制。方法：培养Ishikawa和HEC-1A人子宫内膜癌细胞系,用不同浓度的二甲双胍干预EC细胞。采用划痕试验检测细胞迁徙能力;流式细胞术检测细胞周期和凋亡;Western blot法检测Akt、p-Akt及PTEN的表达。结果：二甲双胍显著抑制Ishika—wa和HEC-1A细胞的迁徙能力（P均〈0．05）。流式细胞术检测显示,二甲双胍使G0／G1期细胞比例显著升高,同时有凋亡诱导作用;Westernblot法检测显示,二甲双胍能下调P—Akt的表达（P均〈0．05）。结论：二甲双胍显著抑制EC细胞的迁徙能力,其作用机制可能与下调p-Akt的表达有关。     

二甲双胍用于子宫内膜癌预防和治疗的研究进展

随着代谢性疾病发病率不断升高,子宫内膜癌的发病率不断增长,发病年龄趋于年轻化。前期研究发现,二甲双胍在多种生殖道恶性肿瘤的预防和治疗中发挥着重要作用。流行病学显示,二甲双胍可降低糖尿病患者子宫内膜癌的发病风险,提高子宫内膜癌患者的总生存率和无瘤生存期。本文就二甲双胍在子宫内膜癌预防和治疗方面的研究进展做一综述。     

二甲双胍治疗子宫内膜癌的研究进展

子宫内膜癌是妇科常见三大恶性肿瘤之一,随着人们生活水平的提高,子宫内膜癌的发病率逐年提升。二甲双胍作为一种常用降糖药,其抗肿瘤作用得到越来越多的关注。二甲双胍可通过PI3K、IGF1、AMPK、p53、caspase、STAT3等多个通路抑制细胞增殖和转移,促进细胞凋亡,提高患者的生存期。二甲双胍和其他药物联用还能减少耐药,协同抗癌。本文现就二甲双胍在子宫内膜癌治疗中的应用及其作用机制的研究现状做一总结。     

人子宫内膜癌耐醋酸甲羟孕酮细胞株的建立

目的建立人子宫内膜癌孕激素耐药细胞株Ishikawa（ISH）/醋酸甲羟孕酮（MPA）,为子宫内膜癌耐孕激素的发病机制提供基础。方法采用逐步递增MPA浓度方法进行Ishikawa细胞株耐药体外诱导。采用MTS法测定药物敏感性;绘制细胞生长曲线和计算群体倍增时间。结果 （1）成功建立了MPA诱导的人子宫内膜癌耐MPA细胞株ISH/MPA,其对MPA耐药指数为3.35;（2）ISH/MPA在含MPA 10μmol/L的培养基中的细胞倍增时间与Ishikawa在普通培养基中的倍增时间比较,差异无统计学意义（P〉0.05）,两者生长曲线比较一致。结论成功建立人子宫内膜癌MPA耐药细胞模型ISH/MPA细胞系,为进一步研究子宫内膜癌耐孕激素机制提供了基础。     

二甲双胍联合化疗药对妇科肿瘤的作用

化疗是治疗恶性肿瘤的主要手段之一,而患者对化疗药的耐药引起的肿瘤复发、恶化是肿瘤治疗中面临的巨大挑战。二甲双胍作为临床中治疗2型糖尿病的一线药物,副作用小、价格低廉。研究表明,二甲双胍联合化疗药能起到更强的抗肿瘤效果,甚至有望逆转肿瘤耐药,这有望成为肿瘤治疗的突破口。本文将对二甲双胍抗肿瘤的作用机制及二甲双胍联合化疗药在妇科肿瘤中的研究进展作一简要综述。     

EGFR过表达降低人子宫内膜癌细胞孕激素敏感性的研究

目的:检测表皮生长因子受体(EGFR)基因在子宫内膜癌孕激素敏感细胞株Ishikawa及孕激素不敏感细胞株KLE的表达,探讨EGFR基因过表达对人子宫内膜癌细胞孕激素敏感性的影响。方法:实时定量PCR法和蛋白印迹法检测Ishikawa和KLE细胞中EGFR及PR-BmRNA和蛋白的表达。将EGFR全长cDNA真核表达质粒在脂质体介导下转染至Ishikawa细胞,同时以转染空载体和未转染的Ishikawa细胞为对照,分别应用实时定量PCR检测各组细胞EGFR、PR-BmRNA表达的变化,应用蛋白印迹法检测各组细胞EGFR、PR-B蛋白表达的变化;CCK-8法观察转染EGFR基因后Ishikawa细胞对孕激素敏感性的变化。结果:(1)Ishikawa细胞中,EGFRmRNA和蛋白的表达明显低于KLE细胞(P<0.001),而PR-BmRNA和蛋白的表达则显著高于KLE细胞(P<0.001);(2)稳定转染EGFR基因后,Ishikawa细胞中EGFRmRNA和蛋白的表达水平明显高于转染空载体和未转染的Ishikawa细胞(P<0.001),而PR-BmRNA和蛋白的表达水平则显著降低;(3)10-8、10-7、10-6、10-5mol/L的MPA对未转染和转染空载体的Ishikawa细胞的抑制作用显著(P<0.05),但对稳定过表达EGFR的Ishikawa细胞无明显抑制作用(P>0.05)。结论:转染EGFR基因能有效提高Ishikawa细胞内EGFR基因的表达,但可下调PR-B基因的表达使Ishikawa细胞对MPA不敏感。     

屈螺酮炔雌醇联合二甲双胍治疗PCOS合并胰岛素抵抗的子宫内膜癌前期及癌早期患者的临床研究

目的:探讨PCOS合并胰岛素抵抗的子宫内膜癌早期及癌前患者采用屈螺酮炔雌醇联合二甲双胍药物转化治疗的疗效。方法:回顾分析2010年至2015年济宁市第一人民医院、汶上县人民医院收治的接受屈螺酮炔雌醇联合二甲双胍治疗的30例胰岛素抵抗型PCOS的子宫内膜癌前及癌早期病变患者的临床资料。其中采用高效孕激素治疗失败者16例,首次确诊者14例。30例患者中,诊刮病理为子宫内膜癌前病变者20例,子宫内膜样腺癌高分化G1级者10例。治疗前对患者进行全面分期评估,治疗后每3个月诊刮以评价疗效,对有生育要求者随访其生育情况。结果:30例(100%)患者治疗3~6个月后均获得完全缓解。2例(6.7%)复发。有生育要求的13例患者中,6例(46.2%)成功妊娠。结论:对PCOS合并胰岛素抵抗要求保留子宫的子宫内膜样腺癌早期及子宫内膜不典型增生的患者,采用屈螺酮炔雌醇联合二甲双胍的治疗效果高于单一孕激素。     

三氧化二砷对子宫内膜癌醋酸甲羟孕酮耐药细胞的作用研究

目的研究三氧化二砷(AS_2O_3)对子宫内膜癌细胞和耐药细胞的作用及机制。方法 (1)分别采用MTS法和Annexin V-FITC/PI双染色流式细胞术检测不同浓度的AS_2O_3在不同作用时间下对ISK细胞增殖和凋亡的影响;(2)采用浓度梯度递增持续刺激诱导法,体外建立MPA耐药细胞后,采用MTS法和Annexin V-FITC/PI双染色流式细胞术,检测AS_2O_3对ISK/MPA增殖和凋亡的影响,并与ISK组对比。(3)Western-blot法检测AS_2O_3作用后ISK与ISK/MPA细胞内p-AKT、p-ERK1/2及Caspase-3、Bcl-2和Bax蛋白的表达变化。(4)建立裸鼠皮下移植瘤模型,腹腔注射2 mg/kg的AS_2O_3,观察裸鼠瘤体的体积变化及毒副作用。结果 (1)AS_2O_3对ISK细胞有生长抑制作用,且呈时间和浓度依赖性;AS_2O_3作用后,ISK的凋亡率呈浓度依赖性升高,48 h细胞凋亡率大于24 h(P0.05);(2)成功建立人子宫内膜癌MPA耐药细胞系;AS_2O_3对ISK/MPA细胞具有生长抑制和促凋亡作用,且呈时间和浓度依赖性;AS_2O_3对ISK细胞组的促凋亡作用强于ISK/MPA细胞组(P0.05),但生长抑制作用比较,差异无统计学意义(P0.05);(3)AS_2O_3作用后,ISK及ISK/MPA细胞内p-AKT、p-ERK1/2和Caspase-3表达降低,Bcl-2表达降低而Bax表达升高(P0.05);(4)成功建立裸鼠皮下移植瘤模型,AS_2O_3作用后,裸鼠瘤体体积明显减小(P0.05)。结论 AS_2O_3对ISK/MPA细胞有增殖抑制和促凋亡作用,机制可能为AS_2O_3可导致Akt、ERK1/2磷酸化水平的降低,抑制P13K/AKT通路和MPAK/ERK通路的激活及在下调Bcl-2表达的同时,上调Bax蛋白的表达,继而调节凋亡相关蛋白通路分子Caspase-3的表达发挥增殖抑制和促凋亡作用。     

子宫内膜癌患者孕激素抵抗机制的研究

子宫内膜癌是发达国家最常见的妇科恶性肿瘤之一,也是女性癌症患者的第八大死亡因素,在我国其发病率仅次于宫颈癌。对早期、复发及有生育要求的子宫内膜癌患者,孕激素治疗是主要治疗措施。但部分子宫内膜癌患者对孕激素治疗不敏感(即孕激素抵抗),给临床治疗带来困难。研究发现,在机体循环过程中有多种因素可导致孕激素抵抗,其中孕激素受体的异常改变是目前孕激素抵抗的主要原因。此外,Fas/Fasl、PI3K/Akt、Survivin、EGFR及胰岛素抵抗等都与孕激素抵抗密切相关。     

二甲双胍辅助治疗妇科肿瘤的研究进展

二甲双胍是临床治疗糖尿病的常用药,具有安全、经济等优势。目前临床前研究及临床试验提出,除降糖作用外,二甲双胍还可以在妇科三大恶性肿瘤的治疗中发挥抗肿瘤的辅助作用。已有研究显示,二甲双胍可以降低子宫颈癌、卵巢癌、子宫内膜癌的复发风险,并提高放化疗药物疗效。具体而言,二甲双胍通过多途径影响肿瘤细胞的生长和代谢,抑制肿瘤的进展。它可抑制肿瘤细胞的增殖,诱导肿瘤细胞凋亡,阻断肿瘤细胞的能量代谢,以及调节肿瘤微环境等。这些作用有助于减少肿瘤细胞的生长和扩散,并提高患者对放化疗药物的敏感性。尽管已有证据支持二甲双胍在妇科恶性肿瘤辅助治疗中的潜在作用,但目前仍缺乏明确的证据证实它可以预防肿瘤的发生和发展。因此,在将二甲双胍作为预防妇科肿瘤的药物方面,仍需要进一步的研究来明确其效果和安全性。     

Dose of progestin in postmenopausal-combined hormone therapy and risk of endometrial cancer

OBJECTIVE: We studied the impact of progestin dose on this risk. The pattern and number of days per month that progestin is given in postmenopausal combined hormone therapy appears to affect endometrial cancer risk. We assessed the impact of progestin dose on this risk. STUDY DESIGN: A population-based, case-control study included 647 cases with endometrial cancer and 1209 controls. RESULTS: Among users of estrogen with medroxyprogesterone acetate (MPA) 10 to 24 days/month, women who took >100 mg/month had an endometrial cancer risk that was equal to that of hormone nonusers (95% CI 0.6-1.7). The corresponding relative risk was 0.8 (95% CI 0.5-1.5) in those who used a lower monthly MPA dose for 10 to 24 days/month. Among users of a continuous combined hormone regimen, the risk of endometrial cancer was low relative to hormone nonusers, regardless of MPA dose. CONCLUSION: Among the combined hormone regimens most commonly used by postmenopausal women today, MPA monthly dose bears little or no relation to endometrial cancer risk.     

A patient with stage 1a endometrial carcinoma in whom a solitary recurrent lesion was detected in the external iliac lymph node after MPA therapy

BACKGROUND: In the treatment of endometrial carcinoma, young patients desire the preservation of the uterus, and therefore hormonal therapy has been administered. CASE REPORT: The patient was a 39-year-old nullipara diagnosed with stage 1a endometrial carcinoma. The patient desired the preservation of the uterus, and oral administration of MPA was prescribed for 18 weeks, which after the cancer tissue disappeared. However, about 1 year and 6 months later, the patient was diagnosed as having recurrent endometrial carcinoma in the left external iliac lymph node. CONCLUSION: In the literature, there is no patient with relapse at another site in the absence of endometrial relapse after MPA therapy for stage 1a endometrial carcinoma, as performed in the present patient.     

Metformin is associated with improved survival in endometrial cancer

Objective

Preclinical evidence suggests that metformin exhibits anti-tumorigenic effects in endometrial cancer. We sought to investigate the association of metformin on endometrial cancer outcomes.

Methods

A multi-institutional IRB-approved retrospective cohort analysis was conducted comparing endometrial cancer patients with diabetes mellitus who used metformin (based on medication review at the time of diagnosis) to those who did not use metformin from 2005 to 2010. Metformin use on treatment related outcomes (TTR: time to recurrence; RFS: recurrence free survival; OS: overall survival) were evaluated using univariate and multivariate modeling.

Results

24% (363/1495) endometrial cancer patients were diabetic, of whom 54% used metformin. Metformin users were younger and heavier than non-users, though nearly all were postmenopausal and obese. 75% of both groups had endometrioid histology. Stage, grade, and adjuvant therapy distributions were similar. Metformin users had improved RFS and OS. Non-metformin users had 1.8 times worse RFS (95% CI: 1.1–2.9, p = 0.02) and 2.3 times worse OS (95% CI: 1.3–4.2, p = 0.005) after adjusting for age, stage, grade, histology and adjuvant treatment. Metformin use was not associated with TTR.

Conclusion

Metformin use was associated with improved RFS and OS but not TTR, most likely due to improving all-cause mortality. Its role in modifying cancer recurrence remains unclear. Prospective studies that capture metformin exposure prior to, during and post endometrial cancer treatment may help define the role of metformin upon cancer specific and overall health outcomes.     

Fertility sparing treatment of complex atypical hyperplasia and low grade endometrial cancer using oral progestin

Objective

Oral progestin is an alternative to hysterectomy for women with complex atypical hyperplasia (CAH) or grade one endometrial cancer (G1EC) who wish fertility preservation. We evaluated treatment efficacy and fertility outcomes in this population.

Methods

Women < 45 y treated with oral progestin for CAH or G1EC were identified from two cancer centers. Data were obtained from medical records and telephone questionnaires. Time until complete response (CR), and from CR until recurrence was censored for patients without events and analyzed for associations with patient and treatment characteristics; cumulative incidence functions were used to estimate event probability over time.

Results

44 patients were identified, 19 (43%) with CAH and 25 (57%) with G1EC. Median age was 36.5 y (26–44). 24 (55%) achieved CR (median time: 5.7 months). Older age was associated with a lower likelihood of CR (HR 0.84, p = 0.0003, 95% CI, 0.8–0.9). CR probability appeared to plateau after 12 months of therapy. Among those with CR, 13 (54%) recurred (median time 3.5 y). 24 patients (55%) underwent hysterectomy; 3 (13%) were upstaged. 11 (25%) underwent fertility treatment with the following outcomes: 6 (55%) no pregnancy, 2 (18%) at least one live infant, and 3 (27%) spontaneous abortion. One achieved a live birth without intervention.

Conclusion

Oral progestin is an effective temporizing fertility-sparing treatment for women with CAH/G1EC. Fertility specialist involvement is recommended due to the low live birth rate without intervention. Progestin therapy should be re-evaluated at 1 year in non-responders due to a low probability of success. Hysterectomy is recommended after childbearing due to a high recurrence rate.     

Predictors of resolution of complex atypical hyperplasia or grade 1 endometrial adenocarcinoma in premenopausal women treated with progestin therapy

Objective

To identify clinical and pathologic predictors of response to progestin treatment in premenopausal women with complex atypical hyperplasia (CAH) and Grade 1 endometrial adenocarcinoma (Grade 1 EA).

Methods

Forty premenopausal patients with Grade 1 EA or CAH who underwent progestin therapy for a minimum of 8 weeks were retrospectively identified. Patient characteristics and histopathologic features of pretreatment and first follow-up endometrial specimens were evaluated as predictors of resolution, defined as absence of hyperplasia or carcinoma.

Results

Kaplan-Meier analysis indicated 63% resolution at 18 months of follow-up. Multivariate classification analysis showed that resolution rates were higher in individuals with a low pre-treatment qualitative abnormal architecture score and a BMI < 35 (Standardized Resolution Ratio (SRR) = 1.48, p = 0.03). The diagnosis of benign endometrium or simple hyperplasia on the first follow-up specimen was highly predictive of resolution (SRR = 2.25, p = 0.002). Resolution rates were lower among subjects with a high pre-treatment qualitative abnormal architecture score (SRR = 0.37, p < 0.03) and lowest in subjects whose first follow-up specimen showed persistent complexity, atypia, or carcinoma with adjacent stromal decidualization (SRR = 0.24, p = 0.002).

Conclusions

Clinical and pathologic parameters can predict response to progestin therapy in premenopausal women with CAH and Grade 1 EA. A low likelihood of resolution is predicted by an unfavorable pre-treatment architectural score and lack of pathological response in the first specimen, despite adjacent stromal decidualization.     

Efficacy of oral or intrauterine device-delivered progestin in patients with complex endometrial hyperplasia with atypia or early endometrial adenocarcinoma: a meta-analysis and systematic review of the literature

Objectives

To investigate the efficacy of progestin treatment to achieve pathological complete response (pCR) in patients with complex atypical endometrial hyperplasia (CAH) or early endometrial adenocarcinoma (EC).

Methods

A systematic search identified 3245 potentially relevant citations. Studies containing less than ten eligible CAH or EC patients in either oral or intrauterine treatment arm were excluded. Only information from patients receiving six or more months of treatment and not receiving other treatments was included. Weighted proportions of patients achieving pCR were calculated using R software.

Results

Twelve studies met the selection criteria. Eleven studies reported treatment of patients with oral (219 patients, 117 with CAH, 102 with grade 1 Stage I EC) and one reported treatment of patients with intrauterine progestin (11 patients with grade 1 Stage IEC). Overall, 74% (95% confidence interval [CI] 65-81%) of patients with CAH and 72% (95% CI 62-80%) of patients with grade 1 Stage I EC achieved a pCR to oral progestin. Disease progression whilst on oral treatment was reported for 6/219 (2.7%), and relapse after initial complete response for 32/159 (20.1%) patients. The weighted mean pCR rate of patients with grade 1 Stage I EC treated with intrauterine progestin from one prospective pilot study and an unpublished retrospective case series from the Queensland Centre of Gynaecologic Oncology (QCGC) was 68% (95% CI 45-86%).

Conclusions

There is a lack of high quality evidence for the efficacy of progestin in CAH or EC. The available evidence however suggests that treatment with oral or intrauterine progestin is similarly effective. The risk of progression during treatment is small but longer follow-up is required. Evidence from prospective controlled clinical trials is warranted to establish how the efficacy of progestin for the treatment of CAH and EC can be improved further.     

Metformin potentiates the effects of paclitaxel in endometrial cancer cells through inhibition of cell proliferation and modulation of the mTOR pathway

Objectives

To examine the effects of combination therapy with metformin and paclitaxel in endometrial cancer cell lines.

Methods

ECC-1 and Ishikawa endometrial cancer cell lines were used. Cell proliferation was assessed after exposure to paclitaxel and metformin. Cell cycle progression was assessed by flow cytometry. hTERT expression was determined by real-time RT-PCR. Western immunoblotting was performed to determine the effect of metformin/paclitaxel on the mTOR pathway.

Results

Paclitaxel inhibited proliferation in a dose-dependent manner in both cell lines with IC₅₀ values of 1-5 nM and 5-10 nM for Ishikawa and ECC-1 cells, respectively. Simultaneous exposure of cells to various doses of paclitaxel in combination with metformin (0.5 mM) resulted in a significant synergistic anti-proliferative effect in both cell lines (Combination Index < 1). Metformin induced G1 arrest in both cell lines. Paclitaxel alone or in combination with metformin resulted in predominantly G2 arrest. Metformin decreased hTERT mRNA expression while paclitaxel alone had no effect on telomerase activity. Metformin stimulated AMPK phosphorylation and decreased phosphorylation of the S6 protein. In contrast, paclitaxel inhibited AMPK phosphorylation in the ECC-1 cell line and induced phosphorylation of S6 in both cell lines. Treatment with metformin and paclitaxel resulted in decreased phosphorylation of S6 in both cell lines but only had an additive effect on AMPK phosphorylation in the ECC-1 cell line.

Conclusions

Metformin potentiates the effects of paclitaxel in endometrial cancer cells through inhibition of cell proliferation and modulation of the mTOR pathway. This combination may be a promising targeted therapy for endometrial cancer.     

子宫内膜增殖症保守治疗的疗效观察

目的:观察并评价不同保守治疗方案对子宫内膜复杂性增生(CH)及非典型增生(AH)的治疗效果、并发症及患者可接受性。方法:前瞻性研究2009年1月至2012年12月在山东大学齐鲁医院门诊及住院收治的50岁以下、病理结果证实为CH的患者116例,将患者随机分为3组,分别接受口服孕激素(A组)、口服避孕药(B组)或LNG-IUS(C组)治疗;AH的患者共90例,将其随机分为2组,分别接受口服孕激素或GnRH-a治疗。每3个月取子宫内膜行病理检查,随访观察患者治疗后的病理缓解率及不良反应。结果:(1)CH患者:治疗6个月时,口服孕激素治疗组、口服避孕药治疗组、LNGIUS组的病理缓解率分别为77.5%、73.7%和94.7%(P=0.039),治疗期间体重增加的发生率依次为42.5%、7.9%和2.6%(P=0.001),突破性出血发生率分别为15.0%、5.2%和26.3%(P=0.023),恶心呕吐症状发生率分别为7.5%、5.2%和2.6%(P=0.624)。(2)AH患者:治疗6个月时,口服孕激素治疗组、GnRH-a治疗组的病理缓解率分别为70.5%和60.9%(P=0.197)。口服孕激素治疗组患者出现的不良反应主要包括体重增加(59.1%)、突破性出血(22.7%)、恶心呕吐(13.6%)及头晕乏力(6.8%)。GnRH-a治疗组患者出现的不良反应主要包括潮热(67.4%)、阴道干燥(34.8%)、精神状态改变(32.6%)、头痛乏力(10.9%)。患者均未出现血栓形成及肝酶升高。结论:对50岁以下的CH患者,可首选宫腔内置入LNG-IUS系统。对AH患者,若无生育要求或手术禁忌证,应首选手术治疗;对于50岁以下选择保守治疗的患者,口服孕激素和GnRH-a治疗的疗效相当。保守治疗需在严格监测下进行,以便及时发现癌变。