Zhihong Ai

Weighted Gene Co-expression Network Analysis in Identification of Endometrial Cancer Prognosis Markers

OBJECTIVE Endometrial cancer (EC) is the most common gynecologic malignancy. Identification of potential biomarkers of EC would be helpful for the detection and monitoring of malignancy, improving clinical outcomes. METHODS The Weighted Gene Co-expression Network Analysis method was used to identify prognostic markers for EC in this study. Moreover, underlying molecular mechanisms were characterized by KEGG pathway enrichment and transcriptional regulation analyses. RESULTS Seven gene co-expression modules were obtained, but only the turquoise module was positively related with EC stage. Among the genes in the turquoise module, COL5A2 (collagen, type V, alpha 2) could be regulated by PBX (pre-B-cell leukemia homeobox 1)1/2 and HOXB1(homeobox B1) transcription factors to be involved in the focal adhesion pathway; CENP-E (centromere protein E, 312kDa) by E2F4 (E2F transcription factor 4, p107/p130-binding); MYCN (v-myc myelocytomatosis viral related oncogene, neuroblastoma derived [avian]) by PAX5 (paired box 5); and BCL-2 (B-cell CLL/ lymphoma 2) and IGFBP-6 (insulin-like growth factor binding protein 6) by GLI1. They were predicted to be associated with EC progression via Hedgehog signaling and other cancer related-pathways. CONCLUSIONS These data on transcriptional regulation may provide a better understanding of molecular mechanisms and clues to potential therapeutic targets in the treatment of EC.

Overexpressed epidermal growth factor receptor (EGFR)‐induced progestin insensitivity in human endometrial carcinoma cells by the EGFR/mitogen‐activated protein kinase signaling pathway

Fertility‐sparing management is an important treatment for young women who have endometrial carcinoma (EC). Many patients with EC exhibit insensitivity or resistance to progestin therapy, and the molecular mechanisms of that insensitivity and resistance have been elusive. Epidermal growth factor receptor (EGFR) overexpression has been observed in EC, but the roles of EGFR in progestin resistance have not been investigated.

Correlation between the overexpression of epidermal growth factor receptor and mesenchymal makers in endometrial carcinoma

Objective The objective of this study was to evaluate the effect of overexpression of epidermal growth factor receptor (EGFR) on the expression of epithelial cell markers (E-cadherin and α-catenin) and mesenchymal cell markers (N-cadherin and vimentin) in endometrial carcinoma. Methods The expression of all 4 markers was evaluated in EGFR overexpressing Ishikawa cells, control Ishikawa cells, and KLE cells using reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. The expression of these 4 markers was also determined in cancerous tissues of patients with endometrial carcinoma using immunohistochemical staining. Results Ishikawa cells transfected with EGFR showed decreased expression of E-cadherin and α-catenin and increased expression of N-cadherin and vimentin compared with control Ishikawa cells (p<0.01 for all). The expression of N-cadherin and vimentin was higher and the expression of E-cadherin and α-catenin was lower in stage II-III than stage I and in grade II-III than grade I endometrial carcinoma tissue (p<0.01 for all). Conclusion Decreased expression of epithelial markers (E-cadherin and α-catenin) and increased expression of mesenchymal markers (N-cadherin and vimentin) were observed in human endometrial carcinoma tissue. These findings correlate with high EGFR expression in cultured endometrial carcinoma cells.

Influence of epidermal growth factor receptor inhibitor AG1478 on epithelial-mesenchymal transition in endometrial carcinoma cells.

Objective To analyze the influence of epidermal growth factor receptor inhibitor AG1478 on the proliferation and epithelial-mesenchymal transition in endometrial carcinoma cells. Materials and Methods The inhibitory effect of different concentrations of AG1478 on the proliferation of endometrial carcinoma cells was detected by tetrazolium-based assay. Western blot was applied to investigate the influence of AG1478 on expressions of epithelium-cadherin, &agr;-catenin, neural cadherin, vimentin, matrix metalloproteinase 9 (MMP9), and MMP2 protein in endometrial carcinoma cells. The influence of AG1478 on migration and invasion of endometrial carcinoma cells was examined by Transwell migration and invasion assay, respectively. Results AG1478 could suppress the proliferation of different endometrial carcinoma cells, and cells transfected with epidermal growth factor receptor were more sensitive (P < 0.05). The expression of an increase in epithelial marker proteins and a decrease in mesenchymal marker proteins, MMP9, MMP2 were observed in endometrial carcinoma cells after AG1478 treated. This effect was more obvious in cells transfected with epidermal growth factor receptor (P < 0.05). The migration and invasion ability of endometrial carcinoma cells were suppressed by AG1478, and Ishikawa cells transfected with epidermal growth factor receptor were demonstrated to be more sensitive to AG1478 (P < 0.05). Conclusions Epidermal growth factor receptor inhibitor AG1478 could effectively inhibit the proliferation and epithelial-mesenchymal transition of endometrial carcinoma cells.

Epidermal growth factor receptor signaling pathway involved in progestin‐resistance of human endometrial carcinoma: In a mouse model

Aim:  The aim of these investigations was to study the role of gefitinib (a specific oral epidermal growth factor receptor‐tyrosine kinase inhibitor) on reversing progestin‐resistance in a human endometrial carcinoma xenograft model.

Proteomic identification of PKM2 and HSPA5 as potential biomarkers for predicting high‐risk endometrial carcinoma

Aim:  Endometrial carcinoma (EC) is a common gynecologic malignancy. EC has a favorable prognosis because it is usually diagnosed at an early stage. However, the recurrence rate is high and the prognosis is poor for high‐risk EC. Identification of new biomarkers for the prediction of high‐risk features will help to guide the treatment and improve the prognosis of patients with EC.

Cholesterol Synthetase DHCR24 Induced by Insulin Aggravates Cancer Invasion and Progesterone Resistance in Endometrial Carcinoma

3β-Hydroxysteroid-Δ24 reductase (DHCR24), the final enzyme of the cholesterol biosynthetic pathway, has been associated with urogenital neoplasms. However, the function of DHCR24 in endometrial cancer (EC) remains largely elusive. Here, we analyzed the expression profile of DHCR24 and the progesterone receptor (PGR) in our tissue microarray of EC (n = 258), the existing EC database in GEO (Gene Expression Omnibus), and TCGA (The Cancer Genome Atlas). We found that DHCR24 was significantly elevated in patients with EC, and that the up-regulation of DHCR24 was associated with advanced clinical stage, histological grading, vascular invasion, lymphatic metastasis, and reduced overall survival. In addition, DHCR24 expression could be induced by insulin though STAT3, which directly binds to the promoter elements of DHCR24, as demonstrated by ChIP-PCR and luciferase assays. Furthermore, genetically silencing DHCR24 inhibited the metastatic ability of endometrial cancer cells and up-regulated PGR expression, which made cells more sensitive to progestin. Taken together, we have demonstrated for the first time the crucial role of the insulin/STAT3/DHCR24/PGR axis in the progression of EC by modulating the metastasis and progesterone response, which could serve as potential therapeutic targets for the treatment of EC with progesterone receptor loss.

Bioinformatics analysis reveals potential candidate drugs for cervical cancer

We sought to explore the mechanisms of cervical carcinoma response to epidermal growth factor (EGF), and then identify biologically active small molecules capable of targeting the sub‐pathways that were dysregulated in cervical cancer cells in the response to EGF.

Peritoneal relative to venous serum biomarker concentrations for diagnosis of ectopic pregnancy

PurposeTo retrospectively analyze the relationships of peritoneal serum relative to venous serum (Rp/v) ratios for human chorionic gonadotropin, CA-125, and creatine kinase to the ectopic pregnancy (EP).MethodsIntra-abdominal fluid and venous blood samples of 118 subjects with suspected EP were obtained for biomarker measurements. Rp/v-hCG >1 was considered indicative of EP, and final diagnosis was based on surgical finding of an ectopic chorionic villous or gynecological ultrasound finding of an intrauterine gestational sac.ResultsRp/v-hCG and Rp/v-CA-125 were both significantly greater for abortive as compared to ruptured EP and for the absence as compared to presence of active bleeding. However, neither ratio differed significantly between ampullary and isthmic EP. Rp/v-hCG and Rp/v-CA-125 correlated negatively with hemoperitoneum volume. Rp/v-hCG exhibited only modest predictive value for rupture. However, with Rp/v-CA-125 as the diagnostic criterion for rupture, sensitivity and specificity were 66.7 and 100%, respectively; in patients initially diagnosed with EP, Rp/v-CA-125 values <22.43 effectively predicted rupture. Rp/v-CK did not exhibit significant diagnostic value.ConclusionsRp/v-hCG values >1 combined with positive culdocentesis test findings reliably indicate the presence of EP. In patients initially diagnosed with EP, Rp/v-CA-125 values <22.43 predict tubal rupture.

SRSF10-mediated IL1RAP alternative splicing regulates cervical cancer oncogenesis via mIL1RAP-NF-κB-CD47 axis

High-risk human papillomavirus oncoproteins E6 and E7 are the major etiological factors of cervical cancer but are insufficient for malignant transformation of cervical cancer. Dysregulated alternative splicing, mainly ascribed to aberrant splicing factor levels and activities, contributes to most cancer hallmarks. However, do E6 and E7 regulate the expression of splicing factors? Does alternative splicing acts as an “accomplice” of E6E7 to promote cervical cancer progression? Here, we identified that the splicing factor SRSF10, which promotes tumorigenesis of cervix, was upregulated by E6E7 via E2F1 transcriptional activation. SRSF10 modulates the alternate terminator of interleukin-1 receptor accessory protein exon 13 to increase production of the membrane form of interleukin-1 receptor accessory protein. SRSF10-mediated mIL1RAP upregulates the expression of the “don’t eat me” signal CD47 to inhibit macrophage phagocytosis by promoting nuclear factor-κB activation, which is pivotal in inflammatory, immune, and tumorigenesis processes. Altogether, these data reveal a close relationship among HPV infection, alternative splicing and tumor immune evasion, and also suggests that the SRSF10-mIL1RAP-CD47 axis could be an attractive therapeutic target for the treatment of cervical cancer.