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Shortening of the 3＇ untranslated region： an important mechanism leading to overexpression of HMGA2 in serous ovarian cancer

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Chinese Medical Journal 2014年第3期127卷 494-499页

作者： He Xiangjun Yang Jing Zhang Qi Cui Heng Zhang Yujun Institute of Clinical Molecular Biology Peking University Peoples Hospital Beijing 100044 China Gynecology Oncology Center Peking University Peoples Hospital Beijing 100044 China

Background Oncofetal protein high-mobility-group AT-hook protein 2 （HMGA2） is reactivated in serous ovarian cancer （SOC） and its overexpression correlates with poor *** explore the mechanism,we investigated whether HMGA2 could avoid microRNA regulation due to gene truncation or 3＇ UTR shortening by alternative *** Real-time reverse transcription polymerase chain reaction （RT-PCR） was used to evaluate the abundance of different regions of HMGA2 mRNA in 46 SOC *** amplification of cDNA 3＇ ends （3＇ RACE） and Southern blotting were used to confirm the shortening of 3＇ untranslated region （UTR）.5＇ RACE and Southern blotting were used to prove the mRNA *** No significant difference in the ratio of the stable coding region to the fragile region was observed between SOC and control normal fallopian tubes,indicating that the HMGA2 gene is not truncated in *** degrees of 3＇ UTR shortening in SOC samples were observed by comparing the abundance of the proximal region and distal region of the HMGA2 3＇ *** ratio of the proximal to the distal region of the 3＇ UTR correlated significantly with expression of the HMGA2 coding region in SOC （r=0.579,P ＜0.01）.Moreover,although the abundance of the HMGA2 coding region varied,all samples,including the very low expressed samples,exhibit relatively high levels of the proximal 3＇ UTR region,suggesting a dynamic decay of HMGA2 mRNA from the 5＇ *** shortening of 3＇ UTR and the decay from the 5＇ end were confirmed by 3＇ RACE,5＇ RACE and subsequent Southern *** Heterogeneous 3＇ UTR lengths render HMGA2 susceptible to different levels of negative regulation by microRNAs,which represents an important mechanism of HMGA2 reactivation in SOC.

关键词： HMGA2 overexpression 3' UTR shortening microRNA regulation serous ovarian cancer

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COPS5 inhibition arrests the proliferation and growth of serous ovarian cancer cells via the elevation of p27 level

COPS5 inhibition arrests the proliferation and growth of ser...

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2017年中国肿瘤标志物学术大会暨第十一届肿瘤标志物青年科学家论坛

作者： Hongqin Zhang Ailing Zhong Miaomiao Chen Renquan Lu Department of Clinical Laboratory Fudan University Shanghai Cancer Center

Background: COPS5 is a pivotal subunit of COP9 signalosome complex. It not only acts as the catalytic center of the complex but also stably exists as a monomer. Aberrant expression of COPS5 is correlated with decreased p27 expression. ovarian cancer is the most lethal gynecological malignancies, however, the role of COPS5 in ovarian cancer remain to be clarified. Methods: The expression of COPS5 was predicted by bioinformatics analysis and confirmed by immunohistochemistry. COPS5 and COPS6 gene were downregulated by RNA interference(RNAi). The biological role of COPS5 in the transduced cells were evaluated by cell proliferation, colony formation, apoptosis and cell cycle analysis. The interactions of COPS5 and p27 were identified by immunoprecipitation and immunofluorescence. The biological effects of the specific COPS5 inhibitor CSN5 i-3 and/or MK2206 were accessed by CCK-8 and flow ***: COPS5 expression was significantly increased in serous ovarian cancer(SOC) cells and tissues compared with those controls. SOC cells with COPS5 depletion were arrested in G1/G0-phase and exhibited a reduced proliferation ability and an increased cytoplasmic p27 expression. Whereas, the cells were stuck at S-phase accompanied with an elevation of nucleus p27 expression after blocking COPS5 by CSN5 i-3 and COPS6 suppression. Furthermore, inhibition of COPS5 resulted in an elevation of Akt expression and sensitized ovarian cancer cells to MK2206. Conclusion: Inhibition of COPS5 resulted in the elevation of p27 expression, as well as the formation of a novel COPS6 containing CSN sub-complex. Meanwhile, COPS5 depletion promoted Akt expression which might be a negative feedback mechanism of p27. COPS5 knockdown enhanced the inhibitory effect of MK2206 on SOC cells proliferation. Suppression of COPS5 and Akt offers a potential strategy and facilitates the development of targeting therapy for serous ovarian cancers.

关键词： COPS5 COPS6 serous ovarian cancer p27 Akt

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Up-regulation of GSTT1 in serous ovarian cancer Associated with Resistance to Taxol/Carboplatin

Up-regulation of GSTT1 in Serous Ovarian Cancer Associated w...

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2021年中国肿瘤标志物学术大会暨第十五届肿瘤标志物青...

作者： Jing Zhang Suhong Xie Lei Zhou Renquan Lu Lin Guo Fudan University Shanghai cancer Center

serous ovarian cancer（SOC） is the most common women cancer and the leading cause of cancer-related mortality among the gynaecological malignancies. Although effective chemotherapeutics combined with surgery are developed for the treatment, the five-year survival rate is unsatisfactory due to chemoresistance. To overcome this shortcoming of chemotherapy, we established taxol and carboplatin resistant SOC cell lines for the understandings of the molecular and cellular mechanisms of chemoresistance. Here, we found that these chemoresistant cell lines showed less viability and proliferation, due to more cells arrested at G0/G1 phase. Glutathione-S-transferases-theta1（GSTT1）was significantly upregulated in these chemoresistant cells, along with other chemoresistant ***, GSTT1 expression was also significantly upregulated in the SOC patient tissues after taxol treatment, indicating this upregulation was physiologically relevant to chemotherapy. Further,suppression of GSTT1 expression by shRNA in SOC cell lines led to more sensitivity to drug treatment, through increasing divided cells and promoting cell death. Moreover, the expression of DNA topoisomerase 1（Topo I） was in synergy with that of GSTT1 in the chemoresistant cells, and GSTT1 can bind to Topo I in vitro, which suggested GSTT1 could function through DNA repair mechanism during chemoresistance. In summary, our data imply that GSTT1 may be a potential biomarker or indicator of drug resistance in serous ovarian cancer.

关键词： GSTT1 serous ovarian cancer Drug resistance Topo I

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Aberrant Alternative Polyadenylation is Responsible for Survivin Up-regulation in ovarian cancer

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Chinese Medical Journal 2016年第10期129卷 1140-1146页

作者： Xiang-Jun He Qi Zhang Li-Ping Ma Na Li Xiao-Hong Chang Yu-Jun Zhang Institute of Clinical Molecular Biology Peking University People's Hospital Beijing 100044 China Gynecology Oncology Center Peking University People's Hospital Beijing 100044 China

Background： Survivin is an oncoprotein silenced in normal mature tissues but reactivated in serous ovarian cancer （SOC）. Although transcriptional activation is assumed for its overexpression, the long 3＇-untranslated region （3＇-UTR） in survivin gene, which contains many alternate polyadenylation （APA） sites, implies a propensity for posttranscriptional control and therefore was the aim of our study. Methods： The abundance of the coding region, the proximal and the distal region of survivin mRNA 3＇-UTR, was evaluated by real-time polymerase chain reaction （PCR） in SOC samples, cell lines, and normal fallopian tube （NFT） tissues. The APA sites were confirmed by rapid amplification ofcDNA 3＇ ends and DNA sequencing. Real-time PCR were used to screen survivin-targeting microRNAs （miRNAs） that were inversely correlated with survivin. The expression of an inversely correlated miRNA was restored by pre-miRNA transfection or induction with a genotoxic agent to test its inhibitory effect on survivin overexpression. Results： Varying degrees of APA were observed in SOC by comparing the abundance of the proximal and the distal region of survivin 3＇-UTR, and changes of 3＇-UTR correlated significantly with survivin expression （r = 0.708, P 〈 0.01）. The main APA sites are proved at 1197 and 1673 of survivin 3＇-UTR by DNA sequencing. Higher level of 3＇-UTR proximal region than coding region was observed in NFT, as well as in SOC and cell lines. Among the survivin-targeting miRNAs, only a few highly expressed miRNAs were inversely correlated with survivin levels, and they mainly targeted the distal part of the 3＇-UTR. However, in ovarian cancer cells, restoration of an inversely correlated miRNA （miR-34c） showed little effect on survivin expression. Conclusions： In NFT tissues, survivin is not transcriptionally silenced but regulate posttranscriptionally. In SOC, aberrant APA leads to the shortening of survivin 3＇-UTR which enables it to escape the negative regulation of miRNAs and

关键词： 3'-Untranslated Region Shortening Alternative Polyadenylation MicroRNA serous ovarian cancer Survivin

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Prognostic Significance of Hormonal Receptor Status of Malignant ovarian Tumors

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Journal of Pharmacy and Pharmacology 2017年第6期5卷 323-339页

作者： I. G. Tkalia V. S. Svintsitsky S. V. Nespryadko N. Yu. Lukianova V. F. Chekhun National cancer Institute Kyiv 03022 Ukraine R. E. Kavetsky Institute of Experimental Pathology NAS of Ukraine Kyiv 03022 Ukraine

The objective of this study is to investigate hormonal receptor status of MOT （malignant ovarian tumor） and to evaluate its clinical and prognostic significance. Retrospective analysis of the case reports of 284 patients with MOT of different histogenesis, stages I-IV, and immunohistochemical study of paraffin-embedded tissues were performed. Hormonal receptor status of tumors with different morphology genesis was studied and hormonal receptor phenotype of serous OC （ovarian cancer） was determined. The analysis of correlation between the expression of steroid hormone receptors （receptors to estrogens （ER）, progesterone （PR） and testosterone （TR）） in ovarian tumors, histological type of tumors and clinical morphological parameters were performed. Overall and relapse-free survival rates of the patients with serous OC depending on the hormonal receptor phenotype of the tumor were assessed. Presence of positive expression of steroid hormone receptors in serous OC （ER-66.4%, PR^53.4%, TR-53.0%）, mucinous OC （ER-88.0%, PR-84.0%, TR-60.0%） and in sex cord stromal tumors （ER-74.1%, PR and TR-77.8%） is proved by correlation of all steroid receptors expression with morphology type of ovarian tumors （ER - r = 0.4; PR - r = 0.4; TR - r = 0.3; p 〈 0.05）. Direct correlation between hormonal receptor phenotype of serous OC and the age period of the patients was established （r = 0.5; p = 0.002）： postmenopausal women patients reported the most increased frequency of serous OC with positive hormonal receptor tumor phenotypes （52.4%）, in particular during their late post-menopausal period （39.0%）. Significantly low overall survival among the patients with positive hormonal receptor phenotype of serous OC was recorded （29.5±3.4%） in comparison with the same score in the patients with negative phenotype of tumors （44.5±3.7%）（p 〈 0.05）. Multifactor analysis of Cox-regression model has defined that positive hormonal receptor phenotype of serous OC increases the risk of disease relapse （H

关键词： Malignant ovarian tumors serous ovarian cancer hormonal receptor status estrogen progesterone testosteronereceptors phenotype of tumor.

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