二甲亚砜对K562／ADM耐药细胞株耐药性的逆转作用

目的：研究二甲亚砜对Ｋ５６２／ＡＤＭ耐药细胞的逆转作用，方法：通过加入二甲亚砜后药细胞的生长情况，逆转倍数的计算，细胞内过氧化物染色的阳性程度以及电镜下观察耐药细胞诱导前后的变化。结果：二甲亚砜诱导Ｋ５６２／ＡＤＭ耐药细胞前后生长情况有显著差异且逆转倍数为４．０Ｐｏｘ染色阳性程度明显递增，电镜下可观察诱导后Ｋ５６２／ＡＤＭ耐药细胞有向成熟分化趋势，结论：二甲亚砜对逆转Ｋ５６２／ＡＤＭ耐药性是有益的     

二甲亚砜对K562/ADM耐药细胞株耐药性的逆转作用

目的：研究二甲亚砜对K562／ADM耐药细胞的逆转作用。方法：通过加入二甲亚砜后耐药细胞的生长情况，逆转倍数的计算，细胞内过氧化物染色的阳性程度以及电镜下观察耐药细胞诱导前后的变化。结果：二甲亚砜诱导K562／ADM耐药细胞前后生长情况有显著差异且逆转借故为4．0，Pox染色阳性程度明显递增，电镜下可观察诱导后K562／ADM耐药细胞有向成熟分化趋势。结论：二甲亚砜对逆转K562／ADM的耐药性是有益的。     

二甲双胍对卵巢癌细胞的抗肿瘤作用

目的 探讨二甲双胍对卵巢癌细胞增殖、周期、凋亡和迁移的影响及其作用机制。方法 选取卵巢癌A2780、CAOV3、SKOV3细胞为研究对象,MTT和克隆形成实验检测二甲双胍对细胞增殖的影响,划痕实验和Transwell实验检测二甲双胍对细胞迁移能力的影响,流式细胞术检测二甲双胍对细胞周期和凋亡的影响,Western blot法检测二甲双胍对细胞AMPK磷酸化、mTOR磷酸化、CXCR4和Wnt/β-catenin蛋白表达的影响。结果 随着二甲双胍浓度增加和作用时间延长,卵巢癌细胞存活率明显下降。A2780、CAOV3及SKOV3细胞48 h的IC50分别为16.36、36.65和43.44 mmol/L。与对照组相比,二甲双胍处理后,细胞克隆形成能力和迁移能力被明显抑制,细胞周期阻滞于G0/G1期,细胞凋亡率增加。随着二甲双胍浓度增加,磷酸化AMPK蛋白表达逐渐增加,磷酸化mTOR、CXCR4、Dvl3、β-catenin、CyclinD1、CDK1蛋白表达逐渐降低。结论 二甲双胍对卵巢癌细胞具有抗肿瘤作用,其机制可能与激活AMPK抑制CXCR4介导的Wnt/β-catenin信号通路有...     

二甲双胍在妇科肿瘤治疗中的研究进展

二甲双胍以其稳定的疗效、安全性及低廉的价格,多年来一直应用于糖尿病的临床治疗。近年来研究显示二甲双胍的应用可降低肿瘤的发病、复发及转移率,长期应用二甲双胍还可改善癌症患者预后,延长患者的生存时间。本文旨在总结二甲双胍在妇科肿瘤领域新近的研究成果,并探讨其对妇科肿瘤的防治作用和其分子机制。     

二甲双胍的抗肿瘤作用

二甲双胍作为一种廉价安全高效的糖尿病药物已应用多年,近年来研究显示其对各种肿瘤细胞都有不同程度的抑制作用。二甲双胍的抗肿瘤机制尚未完全明确,可能与糖代谢及蛋白激酶途径有关,也可能与炎症因子介导肿瘤杀伤有关。二甲双胍对不同肿瘤细胞杀伤效果不同,对其他抗肿瘤治疗也有一些辅助作用。     

二甲双胍通过抑制p38MAPK信号通路逆转卵巢癌细胞对顺铂的耐药性

目的:探讨二甲双胍是否能够逆转卵巢癌细胞对顺铂的耐药性,以及其可能的作用机制。方法:CCK-8法检测二甲双胍作用后卵巢癌细胞顺铂耐药株C13K和CP70细胞对顺铂的耐药性变化,以及转染针对切除修复交叉互补基因1(excision repair cross-complemention 1,ERCC1)的特异性小干扰RNA(small interfering RNA,si RNA)后C13K和CP70细胞耐药性的变化。实时荧光定量PCR法检测不同浓度二甲双胍作用后,C13K和CP70细胞中ERCC1基因表达的变化。蛋白质印迹法检测二甲双胍作用后C13K和CP70细胞中腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)信号通路的激活和p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38MAPK)信号通路的抑制情况,以及联合或不联合AMPK信号通路阻断剂Compound C作用后ERCC1蛋白的表达情况。同时,蛋白质印迹法检测p38MAPK信号通路抑制剂SB203580作用后耐药细胞中ERCC1蛋白表达的变化,以及二甲双胍作用联合p38MAPK si RNA转染前后耐药细胞中ERCC1蛋白的表达变化。结果:二甲双胍能够逆转C13K和CP70细胞对顺铂的耐药性(P<0.05)。二甲双胍作用能够降低C13K和CP70细胞中ERCC1 m RNA和蛋白的表达水平(P<0.05),并激活AMPK信号通路(P<0.05);而联合应用AMPK信号通路阻断剂Compound C可以阻断二甲双胍的这种作用(即ERCC1蛋白表达水平升高)(P<0.01)。p38MAPK信号通路抑制剂SB203580作用后,C13K和CP70细胞中ERCC1蛋白表达水平降低(P<0.05);而二甲双胍作用可降低p38MAPK的磷酸化水平(P<0.01),抑制p38MAPK信号通路。二甲双胍作用转染了p38MAPK si RNA的C13K和CP70细胞后,ERCC1蛋白的表达水平无明显变化(P>0.05)。结论:二甲双胍可能逆转卵巢癌顺铂耐药株对顺铂的耐药性,并且这一作用可能是通过抑制p38MAPK信号通路和降低细胞中ERCC1的表达而实现的。     

二甲双胍抑制胰腺癌干细胞生长的实验研究

摘 要：［目的］ 探讨二甲双胍对胰腺癌干细胞的生长抑制作用。［方法］ 利用无血清培养基的超低粘附培养方法获得干细胞球样胰腺癌PANC1细胞,然后通过定量PCR检测干细胞球样PANC1细胞中CD133及PDX-1的表达情况,验证胰腺癌干细胞;利用干细胞球样PANC1细胞的MTT实验、流式周期检测实验、Annexin-Ⅴ细胞凋亡实验、定量PCR凋亡因子检测实验,研究二甲双胍对干细胞球样PANC1细胞增殖、周期及凋亡的影响;借助干细胞球样PANC1胰腺癌细胞构建鼠移植瘤模型,观察二甲双胍对肿瘤生长的影响。［结果］ 二甲双胍能够明显抑制干细胞球样胰腺癌PANC1细胞增殖,且具有浓度依赖性,最大抑制率为81.3%,半数有效抑制浓度IC50为（11.94±1.43）mmol/L;二甲双胍对干细胞球样PANC1细胞的G0/G1期有阻滞作用;二甲双胍可以诱导干细胞球样PANC1细胞凋亡,与空白对照组比较,二甲双胍组干细胞球样PANC1细胞的Bcl-2 mRNA减少,而Bad、Bax mRNA的表达增加;干细胞球样PANC1细胞接种至裸鼠皮下均能成瘤,且二甲双胍能够明显抑制移植瘤的生长。［结论］ 二甲双胍能够抑制胰腺癌干细胞增殖,阻滞G0/G1期,并诱导胰腺癌干细胞凋亡,发挥抗胰腺癌干细胞生长的作用。     

不同方式诱导人卵巢癌顺铂耐药细胞株的比较

目的 研究不同诱导方式建立的卵巢癌顺铂耐药细胞株的耐药机理。方法 采用大剂量冲击法和浓度梯度递增法建立卵巢癌顺铂耐药细胞株Skov3/CDDP-P和Skov3/CDDP-50。结果 Skov3/CDDP-P和Skov3/CDDP-50的耐药指数分别为3．7和48．6,伴有形态改变、生长减慢、细胞周期改变等。耐药细胞株的细胞内药物浓度降低,与多药耐药相关基因（MDR1）、多药耐药相关蛋白（MRP）和肺耐药蛋白（LRP）的表达增强有关。谷胱甘肽S转移酶（GST－π）的表达无明显变化,拓朴异构酶Ⅱ（TOPOⅡ）活性轻度下降。不同诱导方式存在差异。结论 顺铂耐药涉及多个相关基因的表达,持续诱导方式容易产生耐药。     

二甲双胍抗肿瘤作用机制的研究进展

二甲双胍作为Ⅱ型糖尿病患者的口服治疗药物已有50多年的历史。近年来多项研究发现并证实了二甲双胍在抗肿瘤方面的作用。研究发现二甲双胍能通过调控多种信号通路抑制肿瘤细胞的生长,为肿瘤治疗提供了新的思路。全文对近年来二甲双胍抗肿瘤的作用机制进行综述,希望能为肿瘤患者的治疗带来新的方向。     

Impact of metformin on survival outcome in ovarian cancer: a nationwide population-based cohort study

ObjectiveInvestigation of new drugs (INDs) is a tremendously inefficient process in terms of time and cost. Drug repositioning is another method used to investigate potential new agents in well-known drugs. This study assessed the survival impact of metformin medication on ovarian cancer.MethodsA national sample cohort of the Korean National Health Insurance Service Data was analyzed. Cox proportional hazards regression was used to analyzing hazard ratios (HRs) and 95% confidence intervals (CIs) after adjusting for underlying diseases and medications as confounding factors for overall survival (OS) and cancer-specific survival (CSS).ResultsA total of 866 eligible patients were included from among 1,025,340 cohort participants. Among them, 101 (11.7%) were metformin users. No difference in OS was observed between non-users and users. No difference in OS was observed according to age and Charlson Comorbidity Index. Long-term metformin use (≥720 days) was associated with better OS (adjusted HR=0.244; 95% CI=0.090–0.664; p=0.006). A multivariate Cox proportional hazards model showed that long-term metformin use was an independent favorable prognostic factor for OS (HR=0.193; 95% CI=0.070–0.528; p=0.001) but not for CSS (HR=0.599; 95% CI=0.178–2.017; p=0.408).ConclusionLong-term metformin use reduced all-cause mortality, but not CSS in ovarian cancer. Whether metformin itself reduces deaths because of ovarian cancer requires further investigation.     

Abrogated energy-dependent uptake of cisplatin in a cisplatin-resistant subline of the human ovarian cancer cell line IGROV-1

The parental IGROV-1 human ovarian adenocarcinoma cell line was intermittently exposed to increasing concentrations of cisplatin to obtain resistant sublines. A stable resistant subline with a resistance factor of 8.4 had been developed after 9 months and 28 passages, which was denoted IGROV_CDDP. A high correlation coefficient of 0.97 was found between the log cell survival and the DNA-adduct peak level during the process of resistance development. IGROV_CDDP was strongly cross-resistant to carboplatin and doxorubicin and moderately cross-resistant to etoposide, docetaxel, and topotecan. Only minor resistance against 5-fluorouracil was observed, whereas IGROV_CDDP was not cross-resistant to methotrexate. Intracellular accumulation of cisplatin was 65% lower in IGROV_CDDP as compared with parental IGROV-1 at 37  °C under normal conditions. Coincubation of cisplatin with the Na⁺/K⁺-ATPase inhibitor ouabain resulted in a more pronounced decrease in platinum accumulation in IGROV-1 (44% decrease) than in IGROV_CDDP (26% decrease). Under energy-depleting conditions the accumulation of cisplatin in the parental cell line was approximately 60% lower than that observed under normal (energy [i.e., ATP] rich) culture conditions. In contrast, the accumulation in IGROV_CDDP was not affected by ATP-depletion. There appeared to be no significant difference between the intracellular accumulation of platinum in the resistant and sensitive cells under conditions of energy deprivation or when the uptake was studied at 0  °C. In conclusion, abrogation of energy-dependent accumulation in IGROV_CDDP seems to be a major mechanism of resistance to cisplatin in this cell line. Received: 21 January 1997 / Accepted: 22 July 1997     

上皮性卵巢癌内分泌治疗体外实验研究

目的探讨上皮性卵巢癌内分泌治疗的可能性及其机制.方法用不同剂量(0.1μmol、1μmol、10μmol)的肿瘤内分泌治疗,药物三苯氧胺(TAM)和甲孕酮(MPA),作用于人卵巢上皮癌细胞系3AO和HO-8910,动态观察对活细胞数、PCNA和EGFR的表达及细胞凋亡指数的影响.结果TAM和MPA在不同剂量均可使3AO及HO-8910活细胞数明显减少(P＜0.01),并呈时间依赖性;低浓度(≤1μmol)的TAM对PCNA表达的影响无统计学意义(P＞0.05),而高剂量(10μmol)时可明显降低PCNA表达(P＜0.05).不同剂量MPA均可使PCNA表达明显降低(P＜0.01);TAM和MPA在各浓度均可显著抑制3AO细胞EGFR表达并促进凋亡发生(P＜0.01),且呈剂量依赖性,MPA诱导凋亡程度显著高于TAM(P＜0.01).结论TAM在低剂量时对卵巢癌细胞增殖的抑制较弱,高剂量时可明显抑制卵巢癌细胞增殖并诱导细胞凋亡,但均弱于MPA,提示TAM可用于卵巢癌的内分泌治疗,若治疗无效,仍可用MPA治疗.     

卵巢癌细胞对顺铂耐药机制的研究

目的：探明卵巢癌对顺铂产生耐药的机理。方法：采用顺铂体外诱导法建立卵巢癌耐药细胞株ＨＯ－８９１０/２。ＭＴＴ法测定其耐药倍数和交叉耐药性,原子收法测定细胞内Ｐt浓度,分光光度法测定ＧＳＨ、ＧＳＴ,流式细胞仪分析细胞周期,检测bcl-2和Ｐ－ＧＰ的表达,结果：ＨＯ－８９１０/２耐顺铂是亲代细胞ＨＯ－８９１０的６．６倍,与５－Ｆ Ｕ,ＡＤＲ有交叉耐药性。对照亲代细胞,耐药细胞中ＧＳＨ含量与ＧＳＴ活性明显增高,     

A phase II,open-label,non-randomized,prospective study assessing paclitaxel,carboplatin and metformin in the treatment of advanced stage ovarian carcinoma

ObjectiveThe purpose of this study was to assess the efficacy and tolerability of a paclitaxel, carboplatin and metformin regimen in the first-line treatment of advanced-stage ovarian, fallopian tube, and primary peritoneal carcinoma.MethodsEligible subjects underwent surgery and 6 cycles of neoadjuvant or adjuvant dose-dense intravenous paclitaxel (80 mg/m²), carboplatin (area under the curve 5 or 6 on Day 1), and oral metformin (850 mg daily). Study participants who completed their primary therapy and attained a clinically defined complete or partial response (PR) were treated with a planned 12 cycles of paclitaxel (135 mg/m² every 21 days) and metformin (850 mg twice daily) maintenance therapy.ResultsThirty subjects received a median of 6 cycles (range, 5–6) of primary induction chemotherapy and were eligible for response evaluation; twenty-three patients exhibited a complete response, while 3 study patients obtained a PR (an overall response rate of 86.7%). Grade 3–4 hematological toxicity included neutropenia (43.3%), thrombocytopenia (10%) and anemia (36.7%). There was no incidence of grade 3–4 neuropathy although 15 patients (50%) developed grade ≤2 neurotoxicity. Additionally, we observed grade ≤2 diarrhea in 20 (66.7%) subjects. The median progression-free survival was 21 months (range, 3–52) and overall median survival was 35 months (range, 15–61). The subjects also received an aggregate 103 cycles (median, 12; range, 6–12) of maintenance chemotherapy.ConclusionThe study results suggest that the combination of paclitaxel, carboplatin and metformin is associated with moderate efficacy and a reasonable toxicity profile.     

A STUDY OF MULTI-GENE EXPRESSION IN THE HIGHLY METASTASIZING HUMAN OVARIAN CANCER CELL LINE HO-8910PM AND ITS MOTHER CELL LINE HO-8910

Ｒｅｓｅａｒｃｈｅｓｏｆｔｕｍｏｒｍｏｌｅｃｕｌａｒｂｉｏｌｏｇｙｉｎｒｅｃｅｎｔｙｅａｒｓｈａｖｅｐｒｏｖｅｄｔｈａｔｐｒｏｔｏｏｎｃｏｇｅｎｅａｎｄｔｕｍｏｒｓｕｐｒｅｓｏｒｇｅｎｅｐｌａｙｖｅｒｙｉｍｐｏｒｔａｎｔｒｏｌｅｉｎｔｈｅｐｒｏｃｅ...     

Survivin反义寡核苷酸对人卵巢癌细胞 SKOV3的抑制作用

背景与目的:在大多数肿瘤组织中都发现了 survivin的异常高表达,这说明 survivin在肿瘤发生中具有重要作用.本研究探讨 survivin反义寡核苷酸 (survivin ASODN)对人卵巢癌细胞 SKOV3的抑制作用及其机制.方法:脂质体介导 survivin反义寡核苷酸转染 SKOV3,用 MTT法检测细胞抑制率. Western blot法检测相关基因蛋白表达,应用流式细胞术、梯形 DNA片段分析及 DAPI染色光镜观察细胞凋亡情况.激酶活性检测方法测定半胱氨蛋白水解酶 3( caspase-3)活性的变化.结果:脂质体介导 survivin反义寡核苷酸转染后, SKOV3细胞的生长受到明显抑制, 1 000 ng/ml ASODN转染组的细胞抑制率为( 60.30± 2.95)%,明显高于对照组( P< 0.05).凋亡细胞比率由转染前的 0.65%增加至 32.10% ,SKOV3细胞内 survivin基因蛋白表达下调 26.3%, caspase-3活性为 0.998± 0.001,较对照组显著增高( P< 0.01).结论: Survivin ASODN可通过诱导细胞凋亡抑制细胞生长, 具有明显的抗癌作用.     

紫杉醇对卵巢癌细胞端粒酶活性影响的研究

目的观察紫杉醇对卵巢癌HO-8910细胞端粒酶活性的影响,以了解紫杉醇抗癌的可能机理.方法 1～100nmol/L紫杉醇处理HO-8910细胞后,用MTT法测定卵巢癌细胞的生长抑制率,TRAP-银染法测定端粒酶活性变化.结果 1～100nmol/L紫杉醇对卵巢癌细胞有明显抑制作用,呈时间依赖性及剂量依赖性,紫杉醇处理卵巢癌细胞后,对端粒酶活性的检测结果显示,端粒酶活性下调.结论紫杉醇对卵巢癌细胞具有明显抑制作用,其下调端粒酶活性可能是紫杉醇抗癌作用的重要机理之一.     

丹皮酚对人卵巢癌细胞A2780s增殖的影响

目的:探讨丹皮酚(paeonol,Pae)在体外对人卵巢癌A2780s细胞增殖的影响。方法:将不同浓度的Pae作用于人卵巢癌A2780s细胞,应用MTT法检测Pae对细胞增殖的抑制作用,FCM法检测Pae对细胞周期分布及细胞凋亡率的影响。结果:7.81～250.00mg/LPae对A2780s细胞的增殖均有抑制作用,且随着Pae浓度的增加和作用时间的延长,其增殖抑制作用增强。FCM检测结果显示,31.25～250.00mg/LPae作用48h后,G0/G1期和G2/M期细胞比率下降,S期细胞比率上升;细胞出现凋亡,凋亡率随着Pae浓度的增加而上升。结论:一定浓度的Pae具有抑制人卵巢癌细胞增殖和诱导其凋亡的作用。