The preparation of polymer therapeutics capable of synergistic co-delivery systems for chemotherapeutic drugs and genes remains a tough problem in cancer therapy. Herein, we develop a novel co-delivery system for chemotherapeutic drugs and genes. Paclitaxel (PTX) was covalently grafted onto the cationic copolymer poly(ethylene glycol)-b-poly(L-lysine) (PEG-b-PLL) via a disulfide bond, and then CXCL1 shRNA was loaded on the cationic micelles PEG-b-PLL-SS-PTX by electrostatic interaction to construct PEG-b-PLL-SS-PTX/pDNA complex. The chemical structure of the complex was confirmed by gel permeation chromatography, H-1 NMR and Fourier transform infrared spectroscopy. The binding ability of the cationic micelles and pDNA was detected by gel retardation assay. In vitro gene transfection studies revealed an enhanced gene transfection efficiency due to the co-delivery of PTX and pDNA to ovarian cancer cells. In vitro and in vivo data demonstrated that the co-delivery complex exhibited a synergistic role of chemotherapy and gene therapy and possessed enhanced antitumor effects. The strategy of using drug-loaded cationic micelles as gene delivery carriers is a versatile approach to construct a co-delivery system, which might result in more efficient cancer therapy. (C) 2022 Society of Industrial Chemistry.

The aim of the present study was to investigate the effects of human epididymis protein 4 (HE4) on drug resistance and its underlying mechanisms. The associations among proteins were detected by immunoprecipitation and immunofluorescence assays. Then, stably transfected cell lines CAOV3-HE4-L and CAOV3-A2-L expressing HE4 short hairpin (sh)RNAs and ANXA2 shRNAs, respectively, were constructed. MTT assay, immunocytochemistry, western blotting, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and flow cytometry were employed to examine drug sensitivity, as well as the expression and activity of P-glycoprotein (P-gp). HE4 and P-gp in epithelial ovarian cancer tissues were assessed via immunohistochemistry. MicroRNAs that targeted the P-gp gene, ABCB1, were predicted using bioinformatics methods, and their expression was evaluated by RT-qPCR. The common signaling pathways shared by HE4, ANXA2 and P-gp were selected by Gene Set Enrichment Analysis (GSEA). The interaction of HE4, ANXA2 and P-gp were confirmed. P-gp expression was positively associated with HE4 and ANXA2 expression, respectively. Moreover, it was observed that there was no significant rescue of P-gp expression in CAOV3-A2-L cells following the administration of active HE4 protein. In addition, the expression of HE4 and P-gp in ovarian cancer tissues of drug-resistant patients were higher compared with that of the drug-sensitive group (P<0.05). Furthermore, the results revealed that hsa-miR-129-5p was significantly increased accompanied by decreased HE4 or ANXA2 expression and P-gp expression in CAOV3-HE4-L and CAOV3-A2-L cells. GSEA analyses disclosed that HE4, ANXA2 and P-gp genes were commonly enriched in the signaling pathway involved in regulating the actin cytoskeleton. These results indicated that HE4 promotes P-gp-mediated drug resistance in ovarian cancer cells through the interactions with ANXA2, and the underlying mechanism may be associated with decreased expression of hsa-miR-129-5p and dysregulation of the actin cytoskeleton signaling pathway.

The small leucine-rich proteoglycan (SLRP) family is widely expressed in extracellular matrix and aggravates tumor progression. However, epiphycan (EPYC), as a member of the SLRPs family, its biological function in cancer has not been confirmed. Thus, we aimed to clarify the role of EPYC in progression of ovarian cancer (OC), and further analyze the molecular mechanisms implicated in tumorigenesis. Here, we analyzed the differential expression genes of GSE38734, including 4 matched primary OC and metastatic tissues. We obtained OC RNAseqs data from the Cancer Genome Atlas (TCGA) and analyzed the correlation between EPYC expression and OC staging, pathological grading, etc. The expression of EPYC in OC and normal ovarian tissues was compared in Oncomine website. We used siRNAs to interfere the expression of EPYC in ovarian cancer cell line SKOV3. Scratch test, transwell-matrigel chamber, CCK8 assay were used to detect the changes of SKOV3 migration, invasion and proliferation ability after EPYC was interfered. We used R software to make GO and KEGG analysis of related genes of EPYC. We used the Hitpredict website to predict interacting proteins. The results showed that the expression of EPYC in metastatic ovarian cancer was higher than primary ovarian cancer, and that in primary cancer was higher than normal ovaries. After siRNA interferes with EPYC expression, the migration, invasion and proliferation of SKOV3 cells were weakened. EPYC mainly played a role in ECM organization, and involved in PI3K/Akt, focal adhesion signaling pathways. EPYC might interact with PLCG2 and CRK, and be involved in signal transduction.

目的:检测CD44在原发性输卵管癌组织和正常输卵管组织中的表达情况,分析并探讨其临床表达意义.方法:采用免疫组化SP法检测66例原发性输卵管癌组织和27例正常输卵管组织中CD44的表达情况,分析其与输卵管癌患者临床病理参数及预后的关系.结果:CD44主要定位于细胞膜,亦在细胞质着色,在原发性输卵管癌中CD44的高阳性表达率为68.18％,明显低于正常输卵管组织(88.89％)(P<0.05);CD44的表达与输卵管癌患者各临床病理参数之间并无明显差异性(P>0.05);生存曲线分析显示CD44低阳性表达和FIGO分期晚期的输卵管癌患者总体生存率明显降低(P均<0.05);CD44是影响输卵管癌患者独立预后的风险因素.结论:CD44在原发性输卵管癌组织中表达降低,CD44低表达和FIGO分期均与输卵管癌患者不良预后有关.

Annexin IV (ANXA4) is highly expressed in ovarian clear cell carcinoma (OCCC); however, its underlying molecular mechanism in OCCC remains unknown. The present study aimed to identify the molecule that ANXA4 may act on and to determine its underlying molecular mechanism. Immunohistochemistry, co-immunoprecipitation and western blotting were performed to detect the expression and interaction of ANXA4, and its associated proteins. Furthermore, MTT assay, flow cytometry, western blotting and gene expression profile enrichment analysis were performed to identify the potential role and molecular mechanism of ANXA4 in OCCC. The results demonstrated that ANXA4 and nuclear factor-kappa-light-chain-enhancer of activated B cells (NF-kappa B) p50 nuclear expression levels were significantly higher in OCCC tissues compared with other subtypes of ovarian cancer, such as serous and mucinous. In addition, a significantly positive correlation was observed between ANXA4 and NF-kappa B p50 expression in OCCC; however, the expression levels of mutant p53 and ANXA4 were negatively correlated in a linear manner. These results suggest that ANXA4 and NF-kappa B p50 may be potential independent risk factors for poor prognosis. ANXA4 and NF-kappa B p50 were demonstrated to interact and their expression was co-localized. The cBioPortal database was used to construct a protein-protein interaction network between ANXA4, NF-kappa B p50 and p53, and functional pathway analysis indicated that the genes were predominantly enriched in the cell cycle and during apoptosis. Transfection of the ANXA4 gene increased the expression of NF-kappa B p50, as well as its downstream targets, Cyclin D1 and B-cell lymphoma-2 (Bcl-2). Furthermore, transfection of the ANXA4 gene increased proliferation and decreased apoptosis of OCCC cells. Treatment with the NF-kappa B inhibitor, BAY 11-7082, decreased Cyclin D1 and Bcl-2 expression levels. Collectively, the results of the present study suggest that wild p53 activates ANXA4 transcription, promotes its expression and enhances NF-kappa B p50 and ANXA4 interaction. This in turn activates the NF-kappa B signaling pathway, promotes cell cycle progression and inhibits apoptosis, thus contributing to the malignant progression of OCCC. Thus, ANXA4 and NF-kappa B p50 may be used as prognostic biomarkers, and may be molecular therapeutic targets in OCCC.

[Abstact] Objective:TodetecttheexpressionofannexinA2(ANXA2)infallopiantubecarcinomaandtofurther investigatetherelationshipbetweenannexinA2andclinicopathologicalparametersoffallopiantubecarcinomaandits prognosis.Methods:TheexpressionofANXA2in62casesoffallopiantubecarcinomaand25casesofnormalfallopi﹣antubetissuesweremeasuredbyimmunohistochemistry.TherelationshipbetweentheexpressionofANXA2andclini﹣copathologicalparametersandprognosisofpatientswithfallopiantubecancerwerealsoanalyzed.Results:ANXA2 wasmainlyexpressedinthecellmembrane,alsostainedincytoplasm.ThehighpositiveexpressionrateofANXA2in fallopiantubecarcinomawas50%,whichwassignificantlylowerthanthatofnormalfallopiantubetissue(96%),P＜0.01.Withthedegreeofdifferentiationdecreased,thehighpositiveexpressionrateofANXA2wasalsodecreased (P＜0.05).Theanalysissurvivalcurveshowedthattheoverallsurvivalrateofpatientswithfallopiantubecarcinoma withnegativeANXA2expressionwasshortened(P＜0.05).Conclusion:ANXA2expressionisdecreasedinfallopian tubecarcinomaandisassociatedwithpoorprognosisoffallopiantubecarcinoma.Itcanbeusedforclinicalmonitoring ofpatientswithfallopiantubecarcinoma.

The interaction between human epididymis protein 4 (HE4) and annexin A2 (Annexin A2) has been found in ovarian cancer. However, it is dimness whether the interaction exists in other malignant tumors. Real-time PCR, western blotting and immunocytochemistry were used to detect mRNA and proteins expression. Co-immunoprecipitation and double-labeling immunofluorescence were used to detect the interaction among HE4, ANXA2 and MMP2. MTS assay was used to test cell proliferation. Adhesion test was used to test cell adhesion. Flow cytometry was applied to examine cell cycle. The scratch test and Transwell assay was performed to detect the migration and invasion of various malignant cell lines. Here we show that the overexpression of HE4 and ANXA2 in various malignant cells is a common phenomenon. HE4 and ANXA2 are co-localized in the cytoplasm and membrane of various tumor cells. ES-2 cells which had both high expression of HE4 and ANXA2 were much stronger in proliferation, adhesion, invasion, and migration than other tumor cells. HE4–ANXA2–MMP2 could form a triple protein complex. HE4 could mediate the expression of MMP2 via ANXA2 to promote cell migration progress. The interaction of HE4 and ANXA2 exists in various types of cancer cells. HE4 and ANXA2 can promote the proliferation, adhesion, invasion, and migration of cancer cells. HE4–ANXA2–MMP2 form a protein complex and ANXA2 plays the role of “bridge”. They performed together to promote cell migration.

BACKGROUND:Abnormal activation of the classic Wnt signaling pathway is closely related to the occurrence of epithelial cancers. B-cell lymphoma 9 (BCL9), a transcription factor, is a novel oncogene discovered in the classic Wnt pathway and promotes the occurrence and development of various tumors. Ovarian cancer is the gynecological malignant tumor with the highest mortality because it is difficult to diagnose early, and easy to relapse and metastasis. The expression and role of BCL9 in epithelial ovarian cancer (EOC) have not been studied. Thus, in this research, we aimed to investigate the expression and clinical significance of BCL9 in EOC tissues and its effect on the malignant biological behavior of human ovarian cancer cells.METHODS:We detect the expression of BCL9 in ovarian epithelial tumor tissues and normal ovarian tissues using immunohistochemistry and analyzed the relationship between it and clinicopathological parameters and patient prognosis. The expression of proteins was detected by Western blot. The MTT assay, flow cytometry, the scratch assay, and the transwell assay were used to detect cell proliferation, apoptosis, migration, and invasion, respectively. A total of 374 ovarian cancer tissue samples were collected using TCGA database. A gene set enrichment analysis of BCL9 was performed.RESULTS:BCL9 was overexpressed in EOC tissues. The level of BCL9 expression was correlated with the 5-year progression-free survival rate and overall survival rate in ovarian cancer patients and independently predicted the risk of ovarian cancer recurrence. Low BCL9 expression inhibited proliferation, invasion and migration of EOC cells, decreased MMP2 and MMP9 expression of ES-2 cell line, increased the BAX/BCL2 ratio and promoted apoptosis of EOC cells.CONCLUSION:BCL9 is overexpressed in epithelial ovarian tumors, resulting in a poor prognosis for ovarian cancer patients. Low BCL9 expression can promote ovarian cancer cell apoptosis, inhibit proliferation and migration. BCL9 promotes the development of ovarian cancer.

As a tumor-associated carbohydrate antigen, elevated expression of Lewis y promotes the malignant behaviors of tumor cells. Although our preliminary study showed that the increased expression of Lewis y antigen decreased the expression of cell cycle inhibitor protein p27, the relevant mechanism remains unclear. Autophagy and the ubiquitin-proteasome system are two main ways of intracellular protein degradation, whose abnormal activities are closely associated with progression of malignant tumors. In our present study, we constructed two stable transfected cell lines with high expression of Lewis y antigen, named CAOV3-FUT1 and SKOV3-FUT1. We showed that the proportion of cells at S phase was significantly increased after FUT1 transfection, whereas p27 protein was obviously decreased. The autophagy activity, the levels of ubiquitination, and chymotrypsin-like protease activity were increased remarkably in the transfected cells. Interestingly, Lewis y antigen promoted the degradation of p27 by increasing ubiquitin-proteasome activity. In the vivo studies, Lewis y antigen improved the tumorigenic ability of ovarian cancer cells in nude mice and reduced the expression of p27. These findings suggested that Lewis y antigen activated both the autophagy and ubiquitin-proteasome activity and promoted the degradation of p27 through the ubiquitin-proteasome pathway.

Autophagy is modulated by multiple factors including CD147, but little is know about the effects and mechanism by which the modification of CD147 by Lewis y antigen regulates autophagy of ovarian cancer cell. Here, we reported that Lewis y antigen can promote basic autophagy activity and restrain autophagic cell death in ovarian cancer cells. Furthermore, human whole genome expression profile microarrays and massage pathway analysis revealed that during early stages of autophagy in ovarian cancer cells with highly expressing Lewis y antigen, PI3K/Akt-mTOR activity was reduced, in contrast, the PI3K/Akt-mTOR signaling pathway was activated as the length of amino acid deprivation increased, which inhibited eIF4G2 expression, further decreased the transcription of autophagy-related genes, suppressed autophagic cell death. we also elaborated that co-regulates protein degradation in cells via the ubiquitin-proteasome system and the autophagy-lysosome pathway. These findings suggested that the modification of CD147 by Lewis y antigen enhanced the survival ability by promoting basic autophagy activity and restraining autophagic cell death in ovarian cancer , thus playing an important role in ovarian cancer malignant progression.

Overexpression of Human epididymis protein 4 (HE4) related with a role in ovarian cancer tumorigenesis while little is known about the molecular mechanism alteration by HE4 up regulation. Here we reported that overexpressed HE4 promoted ovarian cancer cells proliferation, invasion and metastasis. Furthermore, human whole genome gene expression profile microarrays revealed that 231 differentially expressed genes (DEGs) were altered in response to HE4, in which MAPK signaling, ECM receptor, cell cycle, steroid biosynthesis pathways were involved. The findings suggested that overexpressed HE4 played an important role in ovarian cancer progression and metastasis and that HE4 has the potential to serve as a novel therapeutic target for ovarian cancer.

Objective:To explore the relationship between expression of annexin A2(ANXA2)and clinicopatho-logical parameters and the clinical significance of annexin A2 by detecting the expression of annexin A2 in different endometrial tissues. Methods:Expression of ANXA2 in 76 endometrial carcinoma,27 atypical hyperplasia endometri-um,16 cases of normal endometrium was detected by immunohistochemistry,and the relationship between ANXA2 ex-pression and clinicopathological parameters in endometrial carcinoma was analyzed. Results:The expression levels of ANXA2 in endometrial carcinoma and atypical hyperplasia(96. 1% ,96. 3% )were significantly higher than normal endometrium(62. 5% )(both P < 0. 05). ANXA2 expression was increased with FIGO stage progression and the de-creasing of differentiation degree. The positive expression rates of ANXA2 in ER( - )and PR( - )group(both were 100. 0% )were significantly higher than that of ER( + )and PR( + )groups(87. 5% ,88. 0% ,respectively, both P < 0. 05). Strong expression rate of ANXA2 was significantly higher in lymph metastasis( + )(93. 8% )than in lymph metastasis( - )group(60. 0% )(P < 0. 05). Higher ANXA2 expression level could predict a poor progno-sis(P < 0. 05). Conclusion:The expression level of ANXA2 was significantly increased in endometrial carcinoma and could predict poor prognosis of endometrial carcinoma patients.

Objective To study the structural relationship between annexin A4 and the Lewis y antigen and compare their expression and significance in ovarian clear cell carcinoma, and to explore how annexin A4 fucose glycosylation effects the interaction between annexin A4 and NF-kB p50, and how it promotes tumour progression of ovarian clear cell carcinoma. Methods Structural relationships between annexin A4 and Lewis y antigen were detected using immunoprecipitation. Annexin A4 and Lewis y antigen expression in various subtypes of ovarian cancer tissues was detected by immunohistochemistry, and the relation between their expression was examined. Any interactions between annexin A4 and NF-kB p50 in ovarian clear cell carcinoma were detected by co-immunoprecipitation. Then looked for changes in expression of Lewis y antigen, annexin A4, NF-kB p50 and a number of downstream related molecules before and after transfection annexin A4 or FUT1, and also analyzed changes in biological processes. Results Lewis y antigen is a part of annexin A4 structure. The expression rate of both annexin A4 and Lewis y antigen was significantly higher in ovarian clear cell carcinoma than in other subtypes of epithelial ovarian cancer, and are associated with the clinical stages, chemotherapy resistance and poor prognostic. The interaction between annexin A4 and NF-kB p50 promoted cell proliferation, adhesion, invasion, metastasis ability and autophagy, and inhibits apoptosis, Lewis y enhanced this interaction. Conclusion Annexin A4 contains Lewis y structure, Lewis y antigen modification of annexin A4 enhances its interaction with NF-kB p50, which promotes ovarian clear cell carcinoma malignancy progression.

Background: It is well-known that the treatment and monitoring methods are limited for advanced stage of endometrial carcinoma. Biological molecules with expression changes during tumor progression become potential therapeutic targets for advanced stage endometrial carcinoma. Annexin A2 (ANXA2) has been reported to be overexpressed in recurrent endometrial carcinoma, and the expression of human epididymis protein 4 (HE4) is upregulated in endometrial carcinoma. What's more, ANXA2 and HE4 interacted in ovarian cancer and promoted the malignant biological behavior. We speculated that their interaction may exist in endometrial carcinoma as well. We evaluated the expression and the correlation relationship of ANXA2 and HE4 in endometrial carcinoma.Methods: The expression of ANXA2 and HE4 protein in 84 endometrial carcinoma, 30 endometrial atypical hyperplasia, and 18 normal endometrial tissue samples were then measured using an immunohistochemical assay in paraffin embedded endometrial tissues. The structural relationship between ANXA2 and HE4 was explored by immunoprecipitation and double immunofluorescent staining.Results: ANXA2 and HE4 co-localized in both endometrial tissues and endometrial carcinoma cells. ANXA2 and HE4 were expressed in 95.2 % and 85.7 % of the the endometrial carcinoma, respectively, which were significantly higher than normal endometrium (55.6 % and 16.7 %, both p < 0.05). The expression of ANXA2 and HE4 was significantly correlated with FIGO stage, degree of differentiation, myometrial invasion, and lymph node metastasis. ANXA2 was an independent risk factor for the prognosis of endometrial carcinoma (p < 0.05, hazard ratio [HR] = 8.004). The expression of ANXA2 and HE4 was positively correlated (Spearman correlation coefficient = 0.228, p < 0.05). HE4 was an independent factor for ANXA2 in multivariate linear regression model (p < 0.05).Conclusion: We revealed the co-localization of ANXA2 and HE4 in endometrial carcinoma. Expression levels of ANXA2 and HE4 were closely related to the malignant biological behavior of endometrial carcinoma, and ANXA2 was an independent risk factor for poor prognosis. The expression of ANXA2 and HE4 can affect each other.

The main aim of this study was to explore the molecular structural relationship between annexin II (ANXA2) and Lewis y antigen by determining their expression patterns and clinical significance in ovarian epithelial carcinoma. The structural relationship between ANXA2 and Lewis y antigen was examined using immunoprecipitation and confocal laser scanning microscopy in two ovarian caner cell lines ES-2 and CaoV-3. We also constracted the stably transfected cell lines with low ANXA2 gene expression in order to detect the expression level between ANXA2 and Lewis y. ANXA2 and Lewis y were detected in tissues from malignant, borderline, benign, and normal ovarian tissues using immunohistochemical analysis. ANXA2 and Lewis y were present in both two ovarian cancer cells and ANXA2 contained Lewis y antigen. Moreover, expression of Lewis y antigen in ANXA2 from cell after transfection was higher than that before. Our immunohistochemistry data revealed significantly higher positive expression rates of ANXA2 in malignant ovarian tissues, compared to benign tumor and normal tissue, similar to Lewis y antigen levels in ovarian cancer. Notably, tissues displaying marked expression of ANXA2 simultaneously expressed high levels of Lewis y antigen. A linear correlation between the expression patterns of ANXA2 and Lewis y antigen was evident. Consistently, double-labeling immunofluorescence experiments illustrated co-localization of ANXA2 and Lewis y antigen within the same area. In conclusions, ANXA2 contains Lewis y antigen. Our results further demonstrate a close correlation between the expression levels of the two antigens, which are significantly high in ovarian cancer.

Background: Estrogen plays an important role in the progression of ovarian cancer in humans. FOXP1 belongs to the forkhead/winged-helix transcription factor family, and previous research indicated that FOXP1 functioned as a tumor suppressor gene. FOXP1 may be similar to FOXA1 and is closely related to steroid hormone receptors, but the relationship between FOXP1 and ER currently remains unclear.Methods: Ovarian tumors (60 malignant cases, 26 borderline cases, and 13 benign cases) and 14 normal ovarian tissues were collected retrospectively. Immunohistochemistry, western blotting and real-time PCR were used to characterize the expression patterns of FOXP1, ER alpha, and ER beta both at the mRNA and protein levels. We also used co-immunoprecipitation and immunofluorescent colocalization to investigate whether a correlation exists between FOXP1 and ER alpha/ER beta in ovarian cancer tissues.Results: The mRNA level for FOXP1 and ER beta in ovarian carcinoma tissues decreased, while the expression level of ER alpha mRNA increased compared with normal ovarian tissues. With an increase in the degree of ovarian carcinoma malignancy, the ER alpha expression level also increased. The expression pattern of ER beta in ovarian neoplasms was similar to that of the FOXP1 protein; presenting nuclear staining decreased, while cytoplasmic expression increased. Colocalization of FOXP1, ER alpha, and ER beta was present in the cytoplasm, with ER beta specific co-localization with FOXP1 in the perinuclear area. While immunoprecipitates created with FOXP1 mouse anti-human monoclonal antibody showed a positive reaction to an anti-ER antibody, irnmunoprecipitates containing anti-ER antibody and react to anti-FOXP1 antibody.Conclusion: Interactions between FOXP1 and ER may play a pivotal role in the progression of ovarian cancer, and the activation or induction of FOXP1 and ER beta expression in cancer cells may inhibit tumor proliferation. (C) 2014 Elsevier B.V. and Societe francaise de biochimie et biologie Moleculaire (SFBBM). All rights reserved.