IGF1和IGF1R及AKT与卵巢癌顺铂耐药相关性研究

目的:检测胰岛素生长因子(IGF)信号通路关键蛋白IGF-1、IGF-1R及Akt在顺铂耐药卵巢癌患者血清中的表达,及其与多药耐药蛋白MRP2的相关性.方法:利用ATP-TCA法对40例卵巢癌标本进行药敏试验,其中16例对顺铂耐药,24例对顺铂敏感.ELISA法分别检测顺铂耐药和敏感组患者血清中IGF-1、IGF-1R及Akt和MRP2的表达,进行相关性分析.结果:IGF-1、IGF-1R及Akt在卵巢癌顺铂耐药组的表达显著高于敏感组,P值均为0.000 1;MRP2的表达卵巢癌顺铂耐药组高于敏感组,P=0.035; IGF-1和IGF-1R有显著相关性,r=0.755,P=0.0001;IGF-1、IGF-1R和Akt有显著相关性,r值分别为0.812和0.643,P值均为0.000 1;MRP2和IGF-1有相关性,r=0.493,P=0.018;IGF-1R与MRP2无相关性,r=0.231,P=0.173; Akt和MRP2无相关性,r=0.274,P=0.106.结论:IGF信号通路关键蛋白IGF-1、IGF-1R及Akt可能参与卵巢癌顺铂耐药,IGF-1与多药耐药蛋白有一定的相关性,IGF-1可能作为卵巢癌靶向治疗的新靶点.     

卵巢癌患者血清中IGF1、IGF1R及AKT表达、血清CA125半衰期与耐药的关系

目的：检测IGF信号通路关键蛋白IGF1、IGF1R及AKT在顺铂耐药卵巢癌患者血清中的表达,检测血清CA125的动态变化,探讨与卵巢癌耐药患者预后的关系。方法：利用ATP-TCA法对40例卵巢癌标本进行药敏试验,其中16例对顺铂耐药,24例对顺铂敏感,ELISA法分别检测顺铂耐药和敏感组患者血清中IGF1、IGF1R及AKT的表达。采用微粒子捕捉免疫发光技术（MEIA）测定40例卵巢上皮癌患者治疗前、手术前及手术后7-14天、每疗程化疗后血清中CA125浓度,计算CA125半衰期,探讨与预后的关系。结果：IGF1、IGF1R及AKT在卵巢癌顺铂耐药组的表达显著高于敏感组（P值均为0.0001）,顺铂耐药患者CA125半衰期与敏感组患者相比有统计学意义。结论：IGF信号通路关键蛋白IGF1、IGF1R及AKT可能参与卵巢癌顺铂耐药,CA125半衰期联合IGF1、IGF1R及AKT检测可早期预测卵巢癌化疗耐药。     

卵巢癌患者血清中 IGF1、IGF1R 及 AKT 表达、血清 CA125 半衰期与耐药的关系

目的:检测IGF信号通路关键蛋白IGF1、IGF1R及AKT在顺铂耐药卵巢癌患者血清中的表达,检测血清CA125的动态变化,探讨与卵巢癌耐药患者预后的关系.方法:利用ATP-TCA法对40例卵巢癌标本进行药敏试验,其中16例对顺铂耐药,24例对顺铂敏感,ELISA法分别检测顺铂耐药和敏感组患者血清中IGF1、IGF1R及AKT的表达.采用微粒子捕捉免疫发光技术(MEIA)测定40例卵巢上皮癌患者治疗前、手术前及手术后7-14天、每疗程化疗后血清中CA125浓度,计算CA125半衰期,探讨与预后的关系.结果:IGF1、IGF1R及AKT在卵巢癌顺铂耐药组的表达显著高于敏感组(P值均为0.0001),顺铂耐药患者CA125半衰期与敏感组患者相比有统计学意义.结论:IGF信号通路关键蛋白IGF1、IGF1R及AKT可能参与卵巢癌顺铂耐药,CA125半衰期联合IGF1、IGF1R及AKT检测可早期预测卵巢癌化疗耐药.     

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。     

IGF信号通路关键蛋白IGF1、IGF1R和AKT在原发性肺腺癌中的表达及意义

目的检测IGF信号通路关键蛋白IGF1，IGF1R和AKT在原发性肺腺癌中的表达，探讨其与临床病理学特征和生存时间的关系。方法采用免疫组化方法和免疫印迹技术检测IGF1，IGF1R和AKT在31例原发性肺腺癌及12例良性肺病变组织中的表达。结果IGF1、IGF1R和AKT在肺腺癌中的表达率分别为41．9％（13／31）、67．7％（21／31）和51．6％（16／31），显著高于良性肺组织（P值分别为0．0252、0．0016和0．0071）。IGF1和IGF1R及IGF1和AKT在肺腺癌中表达呈显著相关（P值分别为0．0344和0．0179）。晚期肺癌（Ⅲ＋Ⅳ）IGF1和IGF1R表达显著高于早期（Ⅰ＋Ⅱ）（P值分别为0．0109和0．0303）。IGF1、IGF1R和AKT在伴有淋巴结转移肺癌中的表达显著高于无淋巴结转移肺癌（P值分别为0．0468、0．0490和0．0443）。低分化肺癌中IGF1和IGF1R表达显著高于中或高分化肺癌（P值分别为0．0484和0．0291）。IGF1和IGF1R阳性患者的生存时间显著短于阴性者（IGF1：10比14个月，P=0．0103；IGF1R：13比26个月，P=0．0056）。IGF1和IGF1R是肺腺癌预后的影响因素，AKT无预后意义。结论IGF信号通路关键蛋白IGF1、IGF1R和AKT表达在肺腺癌的发生和发展中可能起重要作用，进一步的研究有望展示其在肺癌预后和治疗方面的意义。     

人类卵巢癌顺铂耐药细胞株OV1228／cDDP的建立及其生物学特性

 目的　建立人类卵巢癌细胞顺铂耐药细胞株OV1228／cDDP,并对其生物学特性进行检测。方法　采用顺铂浓度梯度递增法,建立人类卵巢癌顺铂耐药细胞株。通过细胞形态学观察、生长曲线和群体倍增时间测定、药物敏感试验、mdr-1和PKC-α基因及蛋白表达水平的测定,来评价OV1228／cDDP的生物学特性。结果　成功建立了OV1228／cDDP耐药细胞株,耐药指数（RI）为5.97,对多柔比星和5-氟尿嘧啶具有一定的交叉耐药性。与OV1228相比,OV1228／cDDP细胞异型性增加、细胞倍增时间延长;mdr-1和PKC-α在基因和蛋白表达水平均明显增高。结论　OV1228／cDDP细胞对顺铂耐药性稳定,并呈现一定的多药耐药特性,为进一步研究耐药逆转途径提供了实验基础。     

TRAP1调控氧化磷酸化在卵巢癌顺铂耐药中的作用研究

目的 探讨肿瘤坏死因子受体相关蛋白1(TRAP1)在顺铂耐药性卵巢癌中的表达以及对顺铂耐药性卵巢癌细胞氧化磷酸化的影响。方法 收集2019-01-10-2020-01-10郑州人民医院手术切除的卵巢癌组织样本41例,其中顺铂敏感26例(顺铂敏感组),顺铂耐药15例(顺铂耐药组),癌旁组织10例(对照组),及细胞株A2780与顺铂耐药株A2780CP,测定上述对象中TRAP1表达。通过转染siRNA沉默A2780CP细胞中TRAP1表达,检测细胞全细胞及线粒体耗氧率(OCR)、ATP产量变化,检测加药顺铂时细胞增殖活性和凋亡率的变化。结果 顺铂耐药组、顺铂敏感组和对照组TRAP1蛋白表达水平分别为1.00±0.30、0.60±0.14和0.22±0.08,差异有统计学意义,F=47.640,P<0.001。A2780CP细胞TRAP1蛋白表达水平(2.24±0.10)高于A2780细胞(1.00±0.05),差异有统计学意义,t=11.383,P=0.001。与A2780细胞相比,A2780CP细胞的顺铂IC₅₀浓度升高,全细胞及线粒体OCR、ATP产量均升高...     

铂类耐药相关基因在卵巢癌顺铂耐药形成过程中表达量的动态变化

目的了解卵巢癌患者在铂类治疗过程中产生获得性耐药相关基因表达动态变化的情况,为临床预后的预测及个体化治疗奠定理论基础。方法采用Benjamini—Hochberg（BH）法对源于TCGA数据库（The Cancer Genome Atlas）的256例敏感和耐药患者的卵巢癌全基因表达谱数据进行差异表达基因筛选。采用DAVID软件中的KEGG模块对筛选出的差异基因进行通路富集,筛选出P〈0．05的通路,对筛选出的通路所包含的基因采用成组t检验找出差异显著的基因（P〈0．05）。对筛选出的基因采用COREMINE工具进行文本挖掘,找出与多药耐药及肿瘤耐药存在线性相关的基因,将其与患者预后进行分析,确定存在差异显著的基因。采用实时荧光定量PCR法检测所筛选出的基因在裸鼠诱导耐药模型不同给药阶段的表达变化情况。结果BH法共筛选出306个差异表达基因,其中上调基因110个,下调基因196个。DAVID软件的KEGG模块分别筛选出5条上调及4条下调通路,成组t检验筛选出有差异的基因共37个,其中上调基因18个,下调基因19个。COREMINE工具检索出与多药耐药及肿瘤耐药存在线性相关的上调基因为ITPA、IMPDH2、RPS7、PDE5A和PDE4D,下调基因为GNAS、CVFR、BUB3、PRKDC、RBL2、SMC1A、CALR、CCNE2及CHEK1。生存分析结果提示CALR及PRKDC基因的表达与患者生存时间有关,CALR和PRKDC基因高表达组的患者生存时间长于低表达组。qRT-PCR检测发现,CALR和PRKDC基因随顺铂注射次数增多,在SKOV3-GFP及SKOV3／DDPii肿瘤组织中的表达量逐渐降低。结论CALR与PRKDC基因可能与卵巢癌铂类耐药及患者的预后密切相关,且随着耐药的产生,CALR和PRKDC基因的表达量逐渐降低。     

卵巢癌细胞中hCTR1的表达与细胞对顺铂耐药的相关研究

目的:探讨人铜离子转运蛋白1(human copper transporter 1,hCTR1)与卵巢癌细胞对铂类药物耐药的关系.方法:构建重组反转录病毒载体pBABEpuro-hCTR1,转染包装细胞后感染人卵巢癌细胞株SKOV-3,用嘌呤霉素筛选得到稳定表达hCTR1的pBABEpuro-hCTR1/ SKOV-3细胞.应用实时荧光定量.PCR(real time fluorogentic quantitative PCR,RFQ-PCR)和Western 印迹法检测pBABEpuro-hCTR1/ SKOV-3细胞中hCTR1 mRNA和蛋白的表达,MTT法检测顺铂(cisplatin, DDP)对pBABEpuro/SKOV-3和pBABEpuro-hCTR1/SKOV-3细胞的增殖抑制率,FCM检测DDP对上述细胞周期的影响,电感耦合等离子质谱法(ion-coupled plasma-mass spectroscopy,ICP-MS)测定细胞内的铂含量.结果:成功构建了hCTR1高表达的SKOV-3稳定细胞株.DDP对pBABEpuro-hCTR1/SKOV-3细胞的IC50值为(18.97±1.24)μmol/L,对pBABEpuro /SKOV-3细胞的IC50值为(29.35±2.12)μmol/L(P<0.01).DDP对pBABEpuro-hCTR1/SKOV-3细胞的S期阻滞作用大于pBABEpuro/SKOV-3细胞.pBABEpuro-hCTR1/SKOV-3细胞对DDP的摄入能力大于pBABEpuro/SKOV-3细胞.结论: hCTR1参与卵巢癌细胞对DDP的转运,其高表达可增强SKOV-3细胞对DDP的摄入能力和敏感性.     

卵巢癌对顺铂耐药机制的研究进展

卵巢癌的发病率在女性生殖器恶性肿瘤中占第三位，但其患者死亡率却居首位。在细胞减灭术的基础上施以以顺铂为主的联合化疗方案已成为卵巢癌的常规治疗方案。据报道：Ⅲ和Ⅳ期的卵巢癌患者采用联合化疗．其完全缓解率可达60％-80％。但在实际临床应用中却并非如此。近30年来卵巢癌患者的5年生存率一直徘徊于30％-50％之间。其主要原因就是卵巢癌对化疗药物产生了耐爱性。约75％-80％的卵巢上皮癌开始对化疗有反应．其余则表现为原发耐药．最终所有化疗患者至少80％出现耐药。因此，耐药的产生直接影响化疗效果及生存率。如何尽早发现耐药度克服耐药是急需解决的问题。     

胰岛素样生长因子-1及其受体与前列腺癌

胰岛素样生长因子-1作为胰岛素样生长因子家族中的重要组成部分,可以通过与胰岛素样生长因子-1受体结合并激活其下游的信号传导通路,发挥广泛的生物学作用,在前列腺癌的发生、发展及转移中亦具有重要的作用。随着研究的深入,针对胰岛素样生长因子-1受体信号通路的特异性靶向药物应运而生,其中,应用最多的是IGF-IR单克隆抗体,在恶性肿瘤包括前列腺癌的治疗中具有广泛的应用价值。     

胰岛素样生长因子-1在乳腺癌组织中的表达

Objective  We investigated the expression of insulin-like growth factor-1 (IGF-1) so as to explore its relationship with carcinogenesis and development of breast cancer. Methods  IGF-1 mRNA levels in tissues of breast cancer, adjacent breast cancer in 70 cases breast cancer patients were analyzed by RT-PCR with the normal breast tissues of paired breast as the control. Results  The level of IGF-1 mRNA expression in breast cancer tissues was significantly higher than that in the paired adjacent to breast cancer tissues, normal mammary gland tissues. The ration of IGF-1/β-actin were 0.679 ± 0.075, 0.463 ± 0.085, 0.305 ± 0.031, respectively. There was significant difference between different groups (P < 0.005). Expression of IGF-1 was associated with lymph node metastasis, pathological staging and estrogen receptor status of breast cancer and no significant relationship with tumor pathological grouping (P > 0.005). Conclusion  The high-level expression of IGF-1 in breast cancer tissues is correlated with carcinogenesis, development and metastasis of breast cancer.     

Insulin-like growth factor-1 receptors and insulin-like growth factor-1-like activity in human primary breast cancer

In the current study the authors have investigated whether human primary breast cancer specimens contain insulin-like growth factor-1 (IGF-1) receptors (IGF-1-R) or IGF-1-like activities. Simultaneously, epidermal growth factor (EGF) receptors (EGF-R) and cytosolic estrogen receptor (ER), progesterone receptor (PR), and EGF-like activity were determined. All tumors assayed contained a single class of specific iodine 125 (125I)-IGF-1 binding sites (Kd: median 106, range 48-755 pM; n = 32) with limited capacity (Bmax: median 147, range 19-11,900 fmol/mg membrane protein). Seventy percent of 44 tumors (50% ER+, PR+), displayed specific 125I-EGF binding with a wide range of values (median 13, range 2-215 fmol/mg protein, n = 31). A positive relationship was apparent between the amount of IGF-1-R with ER and PR (Spearman; 2P less than 0.02 and 2P less than 0.01, respectively; n = 32), whereas for EGF-R a negative relationship was observed (for both 2P less than 0.01; n = 44). All tumors contained endogeneous IGF-1-like and EGF-like activities as measured by radioreceptor assay on acid-ethanol extracted cytosols (median 15, range 3-131 ng/mg protein, n = 78 for IGF-1; and median 86, range 26-517 ng/mg protein, n = 142 for EGF). Tumor contents of IGF-1-like and EGF-like activities showed a negative relationship with ER (2P less than 0.1 for IGF-1, n = 78; and 2P less than 0.001 for EGF, n = 142), and with PR (2P less than 0.05, n = 78; and 2P less than 0.001, n = 142). No relationship was observed between the tumor contents of IGF-1-like and EGF-like activities. In conclusion, these data support the view that IGF-1 and EGF can act as autocrine or paracrine growth factors in human breast cancer.     

Targeting superoxide dismutase 1 to overcome cisplatin resistance in human ovarian cancer

Purpose  Clinical drug resistance to platinum-based chemotherapy is considered a major impediment in the treatment of human ovarian cancer. Multiple pathways associated with drug resistance have been suggested by many previous studies. Over expression of several key proteins involved in DNA repair, drug transport, redox regulation, and apoptosis has been recently reported by our group using a global quantitative proteomic profiling approach. Superoxide dismutase 1 (SOD1) is one of these proteins consistently over-expressed in cisplatin-resistant ovarian cancer cells as compared to their sensitive counterparts, but its precise role in drug resistance is yet to be defined. Method  In the current study, we examined the role of SOD1 in drug resistance by inhibiting its redox activity in cisplatin-resistant ovarian cancer cells using a small-molecule inhibitor, triethylenetetramine (TETA). The effect of TETA was determined by the cell proliferation assay, clonogenic cell survival assay, and SOD1 activity assay. Results  The inhibition of the SOD1 activity enhanced the cisplatin sensitivity in the resistant cells supporting the hypothesis that SOD1 is a key determinant of cisplatin resistance and is an exploitable target to overcome cisplatin drug resistance. Conclusion  SOD1 plays an important role in cisplatin resistance and modulation of its activity may overcome this resistance and ultimately lead to improved clinical outcomes.     

Downregulation of LINC01508 contributes to cisplatin resistance in ovarian cancer via the regulation of the Hippo-YAP pathway

BackgroundSome long non-coding RNAs (lncRNAs) have been found to contribute to cisplatin resistance. Here, we identified a novel lncRNA that was downregulated in cisplatin-resistant to ovarian cancer (OC) cells and aimed to examine the contribution of LINC01508 to cisplatin resistance in OC cells.MethodsDifferences in the lncRNA expression profile between OV2008 and C13K cells were assessed by lncRNA expression microarray. The expression of LINC01508 in ovarian epithelial cells, four OC cells, and OC, benign ovary tumor and normal ovary, cisplatin-resistant and non-resistant OC specimens were evaluated by quantitative real-time polymerase chain reaction (qPCR). The role of LINC01508 in OC cisplatin-resistant was evaluated by cell counting kit-8 (CCK-8), flow cytometry, colony formation, wound healing, Transwell, and tumor growth inhibition study in vivo. The clinical associations of LINC01508 in OC were evaluated using correlation analysis. The effects of verteporfin (VP) on cisplatin were explored to reveal the function of the hippo-YAP pathway on the cisplatin tolerance of C13K.ResultsLINC01508 was downregulated in cisplatin-resistant OC cells and platinum-resistant OC tissue (p<0.01). LINC01508 downregulation was correlated with tumor size, residual tumor, and platinum resistance. The overexpression of LINC01508 improves in vitro and in vivo sensitivity to cisplatin while predicts the poor overall survival which need further follow-up research. The increased level of LINC01508 could suppress the cisplatin resistance of OC cells through the inhibition of the hippo-YAP pathway.ConclusionsThe study proposes that dysregulation of LINC01508 expression results in resistance of OC to cisplatin through the inhibition of the hippo-YAP pathway.     

Effect of lung resistance-related protein on the resistance to cisplatin in human ovarian cancer cell lines

The mechanisms of drug-resistance in human ovarian cancer cells have not been entirely clarified. The purpose of this study was to investigate whether LRP is involved in the resistance of ovarian cancer cell lines to cisplatin and its molecular mechanism. Human ovarian cisplatin-resistant cancer cell lines (A2780/DDP and COC1/DDP) and their parental cisplatin-sensitive cell lines (A2780 and COC1), alone or transfected with antisense LRP-specific oligonucleotides (ODN) or sense ODN, were treated with cisplatin to induce differentiation. Expression of LRP was examined by RT-PCR and Western blot analysis. The sensitivities of cells to cisplatin were assessed using sulforhodamine B (SRB) assay and flow cytometry, and the accumulation and efflux of cisplatin in the cells and isolated nuclei were examined by high performance liquid chromatographic (HPLC) assay. The expressions of LRP in A2780/DDP and COC1/DDP cells were higher than those in A2780 and COC1 cells and conferred resistance to cisplatin. Transfection of LRP AsODN into A2780/DDP and COC1/DDP cells down-regulated LRP expression and reversed the resistance phenotype. Levels of cisplatin accumulating in cells were increased by LRP-specific AsODN and anti-LRP monoclonal antibody. Isolated nuclei from A2780 and COC1 cells or A2780/DDP and COC1/DDP cells incubated with anti-LRP antibody contained more cisplatin than the nuclei of A2780/DDP and COC1/DDP cells not treated with anti-LRP antibody. Efflux of cisplatin was greater from the nuclei of A2780/DDP and COC1/DDP cells than those of A2780 and COC1 cells, and was inhibited by anti-LRP monoclonal antibody. Thus, LRP was involved in the resistance of ovarian cancer cells to cisplatin and has an important role in the transport of cisplatin both in exocytotic vesicles and between the nucleus and cytoplasm.     

p62/SQSTM1 involved in cisplatin resistance in human ovarian cancer cells by clearing ubiquitinated proteins

Mechanisms of cisplatin resistance in cancer cells are not fully understood. Here, we show a critical role for the ubiquitin-binding protein p62/SQSTM1 in cisplatin resistance in human ovarian cancer cells (HOCCs). Specifically, we found that cisplatin-resistant SKOV3/DDP cells express much higher levels of p62 than do cisplatin-sensitive SKOV3 cells. The protein p62 binds ubiquitinated proteins for transport to autophagic degradation, reducing apoptosis induced by endoplasmic reticulum (ER) stress in SKOV3/DDP cells. Knockdown of p62 or inhibition of autophagy using 3-methyladenine resensitises SKOV3/DDP cells to cisplatin. Collectively, our data indicate that p62 acts as a receptor or adaptor for autophagic degradation of ubiquitinated proteins, and plays an important role in preventing ER stress-induced apoptosis, leading to cisplatin resistance in HOCCs.