LINC01503 promotes progression of epithelial ovarian cancer through the miR-342-3p/IGF2R axis / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨LINC01503在上皮性卵巢癌（EOC）中的表达水平和生物学功能及其可能的作用机制。方法：收集2015年5月至2016年5月间在河北医科大学第四医院妇瘤科手术切除并经病理学确诊的85例EOC患者的肿瘤组织和输卵管组织。常规培养人EOC细胞A2780、SKOV3、OVCAR3和OV90及正常人卵巢上皮细胞IOSE80，将si-LINC01503、si-NC及miR-342-3p mimic、miR mimic NC分别转染至SKOV3和A2780细胞，分别作为si-LINC01503组、si-NC组、miR-342-3p mimic组和miR mimic NC组。qPCR法检测EOC组织和细胞中LINC01503的表达水平，Kaplan-Meier法分析LINC01503表达水平与患者生存的关系。双荧光素酶报告基因实验验证LINC01503/miR-342-3p/IGF2R轴相关分子间的靶向关系。平板克隆、划痕愈合和Transwell实验分别检测敲低LINC01503及转染miR-342-3p mimic对A2780和SKOV3细胞增殖、迁移和侵袭能力的影响。WB法检测EOC细胞中LINC01503/miR-342-3p通路对IGF2R蛋白表达的影响。构建A2780细胞裸鼠移植瘤模型，观察敲低LINC01503对移植瘤生长的影响。结果：EOC组织和细胞中LINC01503表达水平分别显著高于输卵管组织和IOSE80细胞（均P<0.01），LINC01503高表达组患者术后PFS和OS均显著短于LINC01503低表达组患者（均P<0.01）。敲低LINC01503、转染miR-342-3p mimic均可抑制EOC细胞的增殖、迁移和侵袭能力（均P<0.01）。敲低LINC01503可下调IGF2R的表达（P<0.01），这一现象可通过转染miR-342-3p inhibitor挽救。敲低LINC01503可抑制A2780细胞裸鼠移植瘤的生长（P<0.01）。结论：在EOC组织和细胞中呈高表达的LINC01503与患者的不良预后密切相关，LINC01503可能通过吸附miR-342-3p影响IGF2R表达进而促进EOC的进展。

Expression and clinical significance of bridging intergrator 1 in epithelial ovarian cancer tissues and cells / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨桥接整合因子1（BIN1）在上皮性卵巢癌（EOC）组织中的表达及其临床意义，以及BIN1对EOC细胞A2780增殖、迁移和侵袭的影响。方法：收集2017年7月至2018年1月河北医科大学第四医院手术切除的67例EOC患者的肿瘤组织及同期因其他妇科疾病手术切除的30例非肿瘤患者的卵巢组织（正常对照组）标本。用免疫组织化学染色法检测EOC组织和非肿瘤卵巢组织中BIN1蛋白的表达水平，χ2检验分析BIN1表达与患者临床病理特征之间的关联，Kaplan-Meier法分析BIN1表达与患者的无病生存期（DFS）和总生存期（OS）之间的关系。用qPCR和WB法检测EOC细胞SKOV3、A2780和人卵巢上皮细胞IOSE80中BIN1 mRNA和蛋白的表达水平。利用基因转染技术将BIN1质粒CMV-MCS-GFP-SV40-Neomycin-BIN1和空载体质粒CMV-MCS-GFP-SV40-Neomycin分别转染到A2780细胞以构建过表达BIN1细胞及其对照，用qPCR和WB法分别检测转染细胞中BIN1 mRNA和蛋白的表达水平，CCK-8、划痕愈合和Transwell实验分别检测过表达BIN1对A2780细胞增殖、迁移和侵袭的影响。结果：EOC组织中BIN1阳性表达率显著低于正常卵巢组织（P<0.01）。BIN1表达与EOC患者较晚的术后病理分期、较差的组织学分级、淋巴结转移及腹膜转移存在正向关联（均P<0.05）；BIN1低表达组患者的DFS和OS均短于BIN1高表达组患者（均P<0.05）。SKOV3和A2780细胞中BIN1 mRNA和蛋白的表达水平均显著低于IOSE80细胞（均P<0.01）；过表达BIN1显著抑制A2780细胞的增殖、迁移和侵袭（P<0.05或P<0.01）。结论：BIN1在EOC组织和细胞中呈低表达状态，与患者的不良预后有关；过表达BIN1可降低EOC细胞A2780的增殖、迁移和侵袭能力。

Expression of lncRNA TOB1-AS1 in epithelial ovarian cancer tissues and its clinical significance / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨ERBB2.1转导蛋白反义RNA1（transducer of ERBB2.1 antisense RNA 1，TOB1-AS1）在上皮性卵巢癌（epithelial ovarian cancer，EOC）组织中的表达情况及其临床意义，初步探讨TOB1-AS1对EOC细胞体外增殖、迁移和侵袭的影响。方法：使用TCGA数据库对EOC组织中TOB1-AS1表达情况进行分析；收集2017年7月至2018年1月在河北医科大学第四医院妇科行肿瘤切除并经病理检查证实为EOC的67例患者的肿瘤组织，收集同期因其他妇科疾病接受手术的30例患者的非肿瘤卵巢组织作为对照。采用qPCR法检测EOC组织和非肿瘤卵巢组织中TOB1-AS1的表达水平，χ2检验分析TOB1-AS1的表达与不同临床病理特征之间的相关性，Kaplan-Meier和Cox比例风险回归模型分析患者生存及预后的潜在影响因素。CCK-8实验、划痕实验和Transwell实验分别检测敲低TOB1-AS1表达对EOC细胞SKOV3和A2780增殖、迁移和侵袭的影响。结果：TCGA数据库中资料和qPCR检测结果均显示，在EOC组织中TOB1-AS1的表达水平显著高于非肿瘤卵巢组织（均P<0. 01）。TOB1-AS1的高表达与EOC患者较晚的FIGO分期、较差的组织分级、淋巴结转移及腹膜转移有关（均P<0.05）。Kaplan-Meier生存分析结果显示，TOB1-AS1高表达组患者术后DFS和OS均短于TOB1-AS1低表达组（均P<0.05）。Cox比例风险回归模型分析结果显示，FIGO分期、淋巴结转移、腹膜转移及TOB1-AS1表达是EOC患者预后的独立影响因素（均P<0.05）。TOB1-AS1在EOC细胞系SKOV3、A2780中的表达水平也显著高于正常卵巢上皮细胞系IOSE80（均P<0.01）。细胞功能实验结果显示，敲低TOB1-AS1可抑制SKOV3和A2780细胞的增殖、迁移和侵袭（均P<0.05）。结论：TOB1-AS1在EOC组织中高表达，与患者的不良预后显著相关。TOB1-AS1可能通过促进EOC细胞SKOV3、A2780的增殖、迁移和侵袭来影响EOC的恶性进展。

Effect of Icaritin on Proliferation， Apoptosis， Migration and Invasion of Human Ovarian Cancer A2780 Cells / 中国实验方剂学杂志

Objective:To study the effect of icaritin on the proliferation， apoptosis， migration and invasion of human epithelial ovarian cancer A2780 cells and the inhibitory mechanism of icaritin against cell invasion and migration via the regulation of epithelial-mesenchymal transformation （EMT）-related molecule expression. Method:A2780 cells were divided into the blank control group and low-， medium-， and high-dose （5， 10， 20 μmol·L^-1） icaritin groups and received the corresponding inventions for 48 h. Cell proliferation and viability were detected using the cell counting kit-8 （CCK-8）. The cellular proliferation inhibition and apoptosis rates were assayed by flow cytometry. The cell invasion and migration were observed in Scratch test and transwell test， followed by the calculation of wound healing rate and migration rate. The protein and mRNA expression levels of EMT-related molecules including E-cadherin， N-cadherin， and Vimentin and tumor invasion and migration-related molecule matrix metalloproteinase-9 （MMP-9） were measured by Western blot and real-time polymerase chain reaction （Real-time PCR）. Result:As revealed by CCK-8 assay and flow cytometry， compared with the blank control group， the icaritin groups all exhibited elevated proliferation inhibition rate （<italic>P</italic><0.01） and apoptosis rate （<italic>P</italic><0.05）. According to the Scratch test and transwell test， compared with the blank control group， the icaritin groups displayed weakened invasion and migration ability and decreased number and rate of cell invasion and migration （<italic>P</italic><0.05）. Western blot and Real-time PCR results showed that the protein and mRNA expression levels of N-cadherin， MMP-9 and Vimentin in each icaritin group were down-regulated as compared with those in the blank control group， while the expression of E-cadherin was up-regulated. Conclusion:Icaritin inhibits the proliferation and promotes the apoptosis of human ovarian cancer A2780 cells， and it inhibits the invasion and migration of A2780 cells possibly by regulating the expression of EMT-related molecules.

Down-regulation of Beclin1 inhibits the resistance of ovarian cancer A2780/DDP cells for the cisplatin / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of Beclin1 knockdown on cisplatin resistance in ovarian cancer A2780 cells and its related mechanisms. Methods: The mRNA and protein expressions of Beclin1 in A2780 cells and drug resistant A2780/DDP cells were determined by qPCR and Western blotting. After transfection with Beclin1 siRNA, the sensitivity of A2780/DDP cells to cisplatin was detected by MTT assay; Cell clone formation and apoptosis were detected by the Colony formation assay and Flow cytometry assay, respectively; cell autophagy was monitored by monodansylcadaverin (MDC) staining. Furthermore, the protein levels of cell autophagy related proteins, lysosomal associated membrane protein Lamp-2 and Cathepsin B were detected by Western blotting. Results: The mRNA and protein expression levels of Beclin1 in cisplatin-resistant A2780/DDP cells were significantly higher than those in A2780 cells (all P<0.05). The expression of Beclin1 was significantly increased in A2780 cells after treated with cisplatin (P<0.05). Beclin1 knockdown promoted cisplatin induced apoptosis of A2780/DDP cells (P<0.05), inhibited autophagy and cell colony formation (all P<0.05), and increased cell sensitivity to cisplatin (P<0.05). Meanwhile, Western blotting showed that Beclin1 knockdown increased the protein levels of cleaved-caspase 3 and Cathepsin B in A2780/DDP cells, while down-regulated the protein expressions of Atg3, Atg7, LC3Ⅱ/Ⅰand Lamp-2 (all P<0.05). Conclusion: Beclin1 knockdown can improve the sensitivity of A2780/DDP cells to cisplatin, and the mechanism may be related to the inhibition of protective autophagy of cells by regulating the expressions of autophagy related proteins, and the regulation of lysosomes, thus further promoting cisplatin-induced apoptosis of drug-resistant cells.

Over-expression of miR-141-3p promotes malignant biological behaviors of ovarian cancer A2780 cells by down-regulating PTEN and activating PI3K/Akt signaling pathway / 中国肿瘤生物治疗杂志

@#Objective: To explore the effect of miR-141-3p on the proliferation, invasion and apoptosis of ovarian cancer cells via targeting PTEN and regulating PI3K/Akt pathway. Methods: Collecting twenty-eight cases pairs of ovarian cancerovarian cancer patients with tumor tissues and adjacent tissues were collected from patients, who from April 2014 to October 2017 were treated in the Department of Obstetrics and Gynecology. qPCR was applied to detect the expression of miR-141-3p in ovarian cancer tissues and cell lines. The relationship between miR-141-3p and PTEN was verified by dual-luciferase reporter gene assay. After over-expression or knockdown of miR-141 and PTEN genes, the cell viability, invasion and apoptosis of ovarian cancer A2780 cells were examined by CCK-8 assay, Transwell assay and Annexin V-FITC/PI double staining flow cytometry assay, respectively. Furthermore, the effect of miR-1413p on PTEN-PI3K/Akt signaling pathway was measured by WB. Results: miR-141-3p is was highly expressed in ovarian cancer tissues and cell lines (P<0.05 or P<0.01). The dual luciferase reporter gene assay confirmed that miR-141-3p targets PTEN was a target of miR-141-3p and downregulates its expression level was down-regulated (P<0.01). Compared with the control group, after knockdown of miR-141-3p, the proliferation ofA2780 cells was significantly inhibited after knockdown of miR-141-3p (at 48 h, 0.36±0.04 vs 0.82± 0.06, P<0.05), and the invasive ability of A2780 cells was significantly reduced (number of transmembrane cells: 215.32±16.04 vs 45.14±7.88, P<0.01), while the apoptotic rate was significantly increased ([1.85±0.26]% vs [9.29±0.65]%, P<0.01). Over-expression of PTEN significantly inhibited the expression of p-Akt and cell proliferation and invasion in A2780 cells (all P<0.01), inhibited cell proliferation and invasion (all P<0.01) and significantly promoted apoptosis (all P<0.01). However, simultaneous over-expression of miR141-3p or addition of IGF-1 wile over-expressing PTEN can offset the above effects. Conclusion: miR-141-3p facilitates the proliferation, invasion and decreases apoptosis of A2780 cells. The mechanism may be related to targeted regulation of PTEN and activation of PI3K/Akt pathway.

Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell line

BACKGROUND: Ovarian cancer is a significant cancer-related cause of death in women worldwide. The most used chemotherapeutic regimen is based on carboplatin (CBDCA). However, CBDCA resistance is the main obstacle to a better prognosis. An in vitro drug-resistant cell model would help in the understanding of molecular mechanisms underlying this drug-resistance phenomenon. The aim of this study was to characterize cellular and molecular changes of induced CBDCA-resistant ovarian cancer cell line A2780. METHODS: The cell selection strategy used in this study was a dose-per-pulse method using a concentration of 100 µM for 2 h. Once 20 cycles of exposure to the drug were completed, the cell cultures showed a resistant phenotype. Then, the ovarian cancer cell line A2780 was grown with 100 µM of CBDCA (CBDCA-resistant cells) or without CBDCA (parental cells). After, a drug sensitivity assay, morphological analyses, cell death assays and a RNA-seq analysis were performed in CBDCA-resistant A2780 cells. RESULTS: Microscopy on both parental and CBDCA-resistant A2780 cells showed similar characteristics in morphology and F-actin distribution within cells. In cell-death assays, parental A2780 cells showed a significant increase in phosphatidylserine translocation and caspase-3/7 cleavage compared to CBDCA-resistant A2780 cells (P < 0.05 and P < 0.005, respectively). Cell viability in parental A2780 cells was significantly decreased compared to CBDCA-resistant A2780 cells (P < 0.0005). The RNA-seq analysis showed 156 differentially expressed genes (DEGs) associated mainly to molecular functions. CONCLUSION: CBDCA-resistant A2780 ovarian cancer cells is a reliable model of CBDCA resistance that shows several DEGs involved in molecular functions such as transmembrane activity, protein binding to cell surface receptor and catalytic activity. Also, we found that the Wnt/3-catenin and integrin signaling pathway are the main metabolic pathway dysregulated in CBDCA-resistant A2780 cells.

Effect of microRNA-134-5p targeting EGFR on growth of ovarian cancer SKOV3 and A2780 cells / 重庆医学

Objective To investigate the effect of microRNA-134-5p (miR-134-5p) targeting epidermal growth factor receptor (EGFR) on the growth of ovarian cancer cells.Methods The ovarian cancer cell lines SKOV3 and A2780 served as the study objects and were divided into the control group (transfecting miR-NC) and experimental group (transfecting miR-134-5p) according to the treatment method.The expression levels of EGFR gene and downstream target protein were detected by qRT-PCR and western blot.The cell cycle distribution and apoptosis were detected by flow cytometry.The proliferation ability of ovarian cancer cells was detected by MTT assay and colony forming assay.Results The expressions of EGFR and downstream target protein in the experimental group were significantly down-regulated.EGFR mRNA in SKOV3 cells was downregulated to 48％ (P＜0.05),and EGFR mRNA in A2780 cells was down-regulated to 47％ (P＜0.05).The cell cycle of cells in the experimental group was significantly inhibited (P＜0.05),and miR-134-5p induced apoptosis through the EGFR target protein (P＜0.05).The proliferation activity and colony forming ability of the experimental group were significantly inhibited (P＜0.05).Conclusion miR-134-5p could promote the cellular cycle arrest and apoptosis,and reduces the proliferation ability of ovarian cancer cells by targetedly inhibiting the EGFR gene.

The inhibitory effect of OSTP-DDP on the growth of ovarian cancer A2780 cells / 中国药理学通报

Aim To study the growth inhibitory effect of the conjugate ( ovarian cancer specific targeting peptide and cisplatin, OSTP-DDP ) that targeting ovarian cancer cells A2780. Methods Using chemical method to syn-thesize OSTP-DDP, ovarian cancer cells A2780 were cul-tured in vitro, using CCK-8 method ( Cell Counting Kit-8) to detect the growth inhibitory effect of ovarian cancer A2780 cells, which were disposed by OSTP-DDP and DDP. Annexin V-FITC was used to detect the cycle and apoptosis effect of ovarian cancer A2780 cells which were disposed by OSTP-DDP and DDP. Results According to the mass spectrometry and the high performance liquid chromatography ( HPLC ) analysis, OSTP-DDP was proved to synthesize successfully. CCK-8 assay showed that both OSTP-DDP and DDP could play the growth in-hibitory effect and showed a concentration-dependent manner when cells were treated in different concentrations (10,20,40,80,160,320μmol·L-1 ) respectively after 24 h, 48 h, 72 h. And the effect of OSTP-DDP was stronger than DDP (P<0. 05), indicated OSTP-DDP had targeted cytostatic effect. The result of the flow cytometry showed that cell cycle was mostly arrested in G1 phase after 72h treated by OSTP-DDP and DDP, the inhibitory effect of OSTP-DDP was stronger than DDP (P<0. 05). The apop-tosis effect of OSTP-DDP was stronger than DDP ( P <0. 01),suggested that OSTP-DDP had a stronger targeting apoptosis-inducing effect. Conclusion OSTP-DDP has the targeting growth inhibitory effect on the ovarian cancer cell A2780, OSTP as a chemotherapeutic drug targeting vector has a great prospect to treat ovarian cancer.

Smilax china L. rhizome extract inhibits nuclear factor-κB and induces apoptosis in ovarian cancer cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the antitumor effects and associated mechanisms of extract of the Smilax china L. rhizome (SCR) on ovarian cancer cells.</p><p><b>METHODS</b>Ovarian cancer cells A2780 were treated with different concentrations of SCR extract (SCRE), and compared with controls. Effects on cell growth were evaluated by cell counting kit-8 (CCK-8) assay; proliferation effects by EdU incorporation assay; cell cycle by propidium iodide staining; apoptosis by annexin V-fluorescein isothiocyanate/propidium iodide; cellular distribution of nuclear factor-κB (NF-κB) by immunofluorescence; protein levels of NF-κB, caspase-3, poly-adenosine diphosphate (ADP)-ribose polymerase (PARP), Bcl-2-associated X protein (Bax), cellular inhibitor of apoptosis (cIAP)-1, anti-X-linked inhibitor of apoptosis protein (XIAP), B-cell lymphoma-extra large (Bcl-XL), B-cell lymphoma-2 (Bcl-2) and AKT by Western blotting; and effects of SCRE combined with cisplatin or adriamycin on A2780 cells by CCK-8 assay.</p><p><b>RESULTS</b>SCRE suppressed A2780 cell proliferation in a dose-dependent manner (P<0.05,P<0.01), arrested cells in G2/M phase and induced apoptosis by activating caspase-3, PARP and Bax. SCRE treatment also correlated with inhibition of NF-κB and downregulation of Bcl-2, Bcl-XL, cIAP-1, XIAP and AKT. SCRE can promote chemosensitivity to cisplatin and adriamycin in A2780 cells (P<0.01).</p><p><b>CONCLUSION</b>SCR effectively inhibits NF-κB, induces apoptosis and reduces chemoresistance to cisplatin and adriamycin in ovarian cancer cells, which might be its molecular basis for treating ovarian cancer.</p>

The analysis of cisplatin resistance in ovarian cancer treatment / 天津医药

Objective To screen the target genes that contribute to cisplatin resistance in ovarian cancer treatment. Methods Gene expression and methylation profiles of ovarian cancer cells that were sensitive or resistant to cisplatin with accession number GSE15709 were downloaded from GEO database. Differential expressed and methylated genes were identi?fied through associating packages in R. DAVID database to screen the enriched GO terms and pathways of the different ex?pressed genes between A2780 and A2780/DDP. Gene Set Enrichment Analysis (GSEA) of different gene was performed against DAVID database. Genes that exhibited difference in both expression and methylation profiles between the two types of ovarian cancer cells as well as genes that present contradictory profile between expression and methylation were verified via qRT-PCR. Results We found 416 different expressed genes and 281 methylated genes between the two types of ovari?an cancer cells respectively. These differential genes were rich in pathways of cell cycle, DNA replication, nucleus division , p53 signaling , and negative regulation of protein modification process etc. Four genes demonstrated contradictory profile be?tween expression and methylation in the two types of ovarian cancer cells and were verified by qRT-PCR. Conclusion Combination of bioinformatics and molecular biology is useful in the identification of target genes that contribute to resis?tance of cisplatin in ovarian cancer treatment and further reveal molecular mechanism behind it.

Determination of free cisplatin in medium by differential pulse polarography after ultrasound and cisplatin treatment of a cancer cell culture.

The in vitro study was carried out for detection of the cisplatin in free form and in culture medium, depending on various conditions of sonodynamic human ovarian cancer cells A2780 treatment by differential pulse polarography (DPP). For sonodynamic treatment, we used cisplatin alone and combined cisplatin/ultrasound treatments. The ultrasound exposure intensity of 1.0 and 2.0 W∙cm-2 in far field for incubation periods 1, 24 and 48 h was used. The parameters of DPP measurements were - 1 s drop time, 5 mV.s-1 voltage scan rate, 50 mV modulation amplitude and negative scanning direction; platinum wire served as counter electrode and Ag|AgCl|3 M KCl as reference electrode. The results showed the dependence of free platinum quantities in culture medium on incubation time and treatment protocol. We found difference in concentration of free cisplatin between conventional application of cisplatin and sonodynamic treatment. The sonodynamic combined treatment of cisplatin and ultrasound field showed a higher cisplatin content in the culture medium than cisplatin treatment alone; a difference of 20% was observed for incubation time 48 h. The results also showed the influence of a time sequence of ultrasound and cytostatics in the sonodynamic treatment. The highest amount of free cisplatin in the solution was found for primary application of cisplatin and the subsequent ultrasound exposure. The quantity of free cisplatin increased with time, namely for time intervals 1-24 h. There was no difference between the DPP signal of cisplatin in reaction mixture containing cells in small quantities and micro-filtered mixture without cells. Thus, the DPP method is suitable for the detection and quantification of free cisplatin in the culture medium of cell suspension. Ultrasound field can be important factor during cytostatic therapy.

Effect of paeonol on proliferation of human ovarian cancer cell line A2780s / 肿瘤

Objective: To investigate the effect of paeonol (Pae) on the proliferation of human ovarian cancer cell line A2780s in vitro. Methods: Inhibitory effects of Pae with different concentrations on A2780s cells were detected by MTT assay. The cell cycle distribution and the apoptosis rate of A2780s cells were determined by flow cytometry (FCM). Results: The proliferation of A2780s cells was significantly inhibited after treatment with Pae (7.81-250.00 mg/L) in a concentration- and time-dependent manner. FCM results showed that the proportion of cells in S phase was increased while which in G0/G1 phase and G2/M phase were decreased after treatment with Pae (31.25-250.00 mg/L) for 48 h. The apoptosis rate of A2780s cells induced by Pae was increased in a concentration-dependent manner. Conclusion: Pae can inhibit the proliferation and induce the apoptosis of human ovarian cancer cells.

Histone Deacetylase Inhibitor Trichostatin A Modulates Cell Cycle in A2780 Human Ovarian Cancer Cell Line / 中国医科大学学报

Objective To analyze the effect and mechanism of trichostatin A（TSA）on cell cycle in human ovarian cancer cells.Methods Human ovarian cancer cell line A2780 cells were cultured in RPMI 1640 supplement.Flow cytometry analysis and RT-PCR were used to examine the distribution of cell cycles and the level of p21WAF/CIPI mRNA.Results TSA induced increase of G2/M cells increased after the treatment of TSA for 36 hours（P 0.05）;the level of p21WAF/CIPI mRNA expression was upregulated after TSA treatment for 12 hours,the highest leve of its expression occurred at 24 hours,the expression level begun to decrease at 48 hours（P 0.05）.TSA simultaneously induced the decrease of S phase cells in a concentration-dependent manne（rP 0.05）.TSA upregulated the expression of p21WAF/CIPI mRNA in a concentration-dependent manner（P 0.05）.Conclusion TSA could block the G2/M phase and inhibits cell proliferation of A2780 cells through upregulating the expression of p21WAF/CIPI mRNA and the activate cyclin-dependent kinase.

Study of xenograft in nude mice on ovarian cancer cell targeted CD59 by siRNA / 中国免疫学杂志

Objective:To investigate the inhibitiory effect of CD59-siRNA on CD59 gene expression and the growth of xenografted ovarian cancer in vivo,and to explore the role of CD59 in tumor immune escape.Methods:A2780 cells transfected with siRNA plasmid (T group),A2780 cells transfected with blank plasmid (V group) and the wild A2780 cells (C group) were subcutaneously injected into nude mice,respectively.The inhibitiory effect of CD59-siRNA on tumor growth and CD59 expression was evaluated by tumor growth curves and in situ hybridization (ISH) and immunohistochemistry (IHC).Results:The tumor growth curves demonstrated that the growth of cells transfected with siRNA plasmid was significantly inhibited (P<0.05).ISH and IHC showed that the expression of CD59 mRNA and CD59 protein were decreased significantly compared with other groups (P<0.05).Conclusion:Studies of in vivo experiment demonstrate that CD59-siRNA significantly inhibites the expression of CD59,increases the sensitivity of A2780 cells to the complement attack and inhibites the tumor growth.The results may further indicate the role of CD59 in tumor immune escape.

MDR1 and MDR3 Genes and Drug Resistance to Cisplatin of Ovarian Cancer Cells / 华中科技大学学报（医学）（英德文版）

To investigate the relationship between MDR1 and MDR3 gene and drug resistance to cisplatin of ovarian cancer cells. Two siRNAs (MDR1, MDR3) which specifically targeted MDR1 and MDR3 genes were transfered into A2780/DDP cells. Then double staining with Annexin- V-FITC/PI was used to detect cell apoptosis by the flow cytometry (FCM). A2780/DDP cell viability was determined by MTT. MDR1 and MDR3 mRNA were assessed by RT-PCR. Caspase-3 protein was detected by Western blotting. Transfection of MDR1 and MDR3 siRNA into A2780/DDP cells failed to reverse the drug-resistance of A2780/DDP cells to cisplatin (P0.05). No significant differ- ence in the apoptosis efficiency was observed between the MDR1 and MDR3 siRNA, pSuppressor- Neo vector transfection cells and untreated cells (P0.05). In the presence of cisplatin of different concentrations, the viability of A2780/DDP cells was not significantly decreased after the transfection. No changes in MDR1 and MDR3 mRNA were found in MDR1 and MDR3 siRNA-transfected A2780/DDP cells. As compared with pSuppressorNeo and untreated groups, no significant difference existed in the expression of MDR1 and MDR3 mRNA (P0.05). The expression of caspase-3 protein in MDR1 and MDR3 siRNA transfected A2780/DDP cells was not significantly increased. It is con- cluded that multidrug resistance induced by cisplatin in ovarian carcinoma cell lines is not due to overexpression of MDR1 and MDR3 gene. The drug resistance of ovarian carcinoma cells to cisplatin is not mediated by P-glycoprotein.

A Study of the Reverse Effect of Tetramethylpyrazine on the Resistant Cell Strain A2780/ADM of Human Ovarian Carcinoma / 医学研究杂志

Objective To study the reverse roles of reverse effect of tetramethylpyrazine on the resistance to adriamycin in human ovarian carcinoma cell line A2780/ADM.Methods The drug sensitivity of A2780/ADM cells was measured by MTT assay.The intracellular concentration of ADM was determined by fluorescent spectrophotometry.The changes of cell apoptosis percentage of A2780/ADM was demonstrated by flow cytometry.Results The IC50 value of TMP in A2780/ADM cell line was significantly decreased by the non-cytotoxic dose or low-cytotoxic dose of TMP.The resistance reversing effect comparing with the IC50 value of ADM cells was respectively 2.07 folds.The intracellular ADM concentration in A2780/ADM cells was significantly elevated by TMP at the same dose and the cell apoptosis percentage was significantly increased by TMP.The expression of P-gp was reduced significantly by TMP.Conclusion TMP could reverse the resistance of human ovarian carcinoma of A2780/ADM cell line to ADM,which may be related to the increased intracellular.