Aberrant metabolic regulation has been observed in human cancers, but the corresponding regulation in human papillomavirus (HPV) infection-associated cervical cancer is not well understood. Here, we explored potential biomarkers for the early prediction of cervical carcinoma based on the metabolic profile of uterine cervical tissue specimens that were positive for HPV16 infection. Fifty-two fresh cervical tissues were collected from women confirmed to have cervical squamous cell carcinoma (SCC; n = 21) or cervical intraepithelial neoplasia (CIN) stages II-III (n = 20). Eleven healthy women constituted the controls (negative controls [NCs]). Real-time polymerase chain reaction (PCR) was performed to detect HPV infection in the tissues. High-resolution magic angle spinning nuclear magnetic resonance was utilized for the analysis of the metabolic profile in the tissues. The expression of rate-limiting enzymes involved in key metabolic pathways was detected by reverse-transcription quantitative PCR. An independent immunohistochemical analysis was performed using 123 cases of paraffin-embedded cervical specimens. A profile of 17 small molecular metabolites that showed differential expression in HPV16-positive cervical SCC or CIN II-III compared with HPV-negative NC group was identified. According to the profile, the levels of α-and β-glucose decreased, those of lactate and low-density lipoproteins increased, and the expression of multiple amino acids was altered. Significantly increased transcript and protein levels of glycogen synthase kinase 3 beta (GSK3β) and glutamate decarboxylase 1 (GAD1) and decreased transcript and protein levels of pyruvate kinase muscle isozyme 2 (PKM2) and carnitine palmitoyltransferase 1A (CPT1A) were observed in the patient group (p < 0.05). HPV infection and cervical carcinogenesis drive metabolic modifications that might be associated with the aberrant regulation of enzymes related to metabolic pathways.

Dipeptidyl peptidase III (DPP3), a zinc-dependent metallopeptidase, is upregulated in a variety of malignancies. However, little is known about its roles in the pathogenesis of these malignancies. The present study was designed to investigate the roles of DPP3 in the pathogenesis and progression of oesophageal cancer (EC). The expression level of DPP3 in EC tissues and adjacent normal tissues was detected in 93 cases of tissue biopsies collected from patients diagnosed with oesophageal carcinoma by immunohistochemistry. The effect of DPP3 expression on cell proliferation, migration or apoptosis was determined in DPP3-depleted EC cells created by infection with lentivirus containing short hairpin RNA specific to the human DPP3 mRNA sequence, followed by detection at the cellular level using a Celigo cell count assay, flow cytometry, wound-healing assay and Transwell assay as well as chip screening with a Human Apoptosis Antibody Array kit, which enables the quantitative detection of 43 apoptosis-related genes. A xenograft model was applied to detect the tumour growth and invasion of DPP3-depleted cancer cells in nude mice. The results revealed that DPP3 expression was elevated in EC tissues compared with adjacent non-tumour tissues, and high DPP3 expression was significantly associated with poor prognosis. DPP3 depletion resulted in reduced cell proliferation and migration and enhanced cell cycle arrest and apoptosis of EC cells and led to the inhibition of tumour growth and invasion in a xenograft model. In addition, DPP3 depletion was associated with the upregulation of the proapoptotic proteins SMAC and p53 and the downregulation of the antiapoptotic proteins clAP-2, IGFBP-2 and TRAILR-4. Finally, DPP3 may promote cell proliferation, migration and survival of EC cells in vitro and tumour growth and invasion of oesophageal carcinoma in vivo, and thus may serve as a molecular target for tumour therapy.

Expression of matrix metalloproteinase-9 (MMP-9) in different degrees of chronic hepatitis B (CHB) and the correlation of MMP-9 with inflammation was investigated. A total of 96 CHB patients (observation group) admitted and treated in Dongying People's Hospital from December 2016 to November 2017 were selected, and they were compared with 60 healthy individuals (control group) selected in the physical examination center during the same time period. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of MMP-9, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), MMP-9 expression in different inflammation grades and fibrosis stages, and the relationship between MMP-9 and inflammation was analyzed. The levels of MMP-9, TNF-α and IL-6 in serum in the observation group were obviously higher than those in the control group (P<0.05). The rank sum test indicated that there were statistically significant differences in the expression levels of MMP-9 among the inflammation grades G0, G1, G2, G3 and G4 (P<0.05). The expression levels of MMP-9 in fibrosis stages S0, S1, S2, S3 and S4 were detected via the rank sum test, and it suggested that the differences among the 5 stages were statistically significant (P<0.05). Pearsons correlation analysis showed that the MMP-9 expression level was positively correlated with TNF-α and IL-6 (P<0.05). In conclusion, the MMP-9 level is elevated remarkably in patients with varying degrees of CHB, it may play an important role in the pathological progress of liver, and it has a close correlation with inflammation, which can provide a theoretical basis for clinical treatment.

Cervical cancer (CC), an aggressive form of squamous cell carcinoma, is characterized by early-stage lymph node metastasis and an extremely poor prognosis. The authors have previously demonstrated that patients with CC have aberrant glycolysis. The upregulation of receptor for activated C kinase 1 (RACK1) is associated with CC lymph node metastasis (LNM). However, its role in mediating aerobic glycolysis in CC LNM remains unclear. In the present study, 1 H nuclear magnetic resonance analysis revealed a significant association between RACK1 expression and the glycolysis/gluconeogenesis pathway. Additionally, RACK1 knockdown inhibited aerobic glycolysis and lymphangiogenesis in vitro and suppressed CC LNM in vivo. Furthermore, protein kinase B (AKT)/ mammalian target of rapamycin (mTOR) signaling was identified as a critical RACK1-regulated pathway that increased lymphangiogenesis in CC. Co-immunoprecipitation, immunofluorescence and western blot analysis revealed that RACK1 activated AKT/mTOR signaling by interacting with insulin-like growth factor 1 receptor (IGF1R). POU class 2 homeobox 2 (POU2F2) bound to the RACK1 promoter and regulated its transcription, thereby functionally contributing to glycolysis and lymphangiogenesis in CC. Of note, the administration of 2-deoxy-D-glucose, which attenuates glycolysis, inhibited RACK1-induced lymphangiogenesis in CC. The correlations between RACK1, IGF1R, POU2F2 and hexokinase 2 were further confirmed in CC tissues. Thus, RACK1 plays a crucial role in CC tumor LNM by regulating glycolysis via IGF1R/AKT/mTOR signaling. Thus, the targeting of the POU2F2/RACK1/IGF1R/AKT/mTOR signaling pathway may provide a novel treatment strategy for CC.

Background: Dipeptidyl peptidase III (DPP3) is a zinc-dependent metallopeptidase and elevated in a variety of malignant tumors, but the underlying mechanism is not well understood so far. Here we investigated the association of esophageal carcinogenesis with the regulation of DPP3 expression by tissue-based quantitative analysis and the depletion of DPP3 expression in esophageal cancer cells and xenograft model. Methods: The expression level of DPP3 in esophageal cancer tissues and adjacent normal tissues was detected in 93 cases of tissue biopsies collected from patients diagnosed with esophageal carcinoma by immunohistochemistry. The effect of DPP3 expression on cell proliferation, migration or apoptosis was determined in DPP3-depleted esophageal cancer cells created by infection with the lentivirus containing the shRNA specific to human DPP3 mRNA sequence followed by cytometric detection using celigo cell count assay, flow cytometry, wound-healing assay and trans-well assay as well as chip screening with a Human Apoptosis Antibody Array kit, which enables the quantitative detection of 43 apoptosis-related genes. A xenograft model was applied to the detection of tumor growth and invasion of DPP3-depleted cancer cells in nude mice.Results: DPP3 expression was elevated in esophageal cancer tissues compared with adjacent non-tumor tissues (normal controls) with statistical significance (P<0.05), and associated with poor prognosis of esophageal carcinoma. The DPP3-depletion resulted in a reduced cell proliferation and migration and enhanced cell-cycle arrest and apoptosis of esophageal cancer cells, and lead to the inhibition of tumor growth and invasion in xenograft model. In addition, DPP3-depletion was associated with the upregulation of pro-apoptotic proteins and the downregulation of anti-apoptotic proteins.Conclusions: These findings suggest that DPP3 may promote cell proliferation, migration and survival of esophageal cancer cells in vitro, and tumor growth and invasion of esophageal carcinoma in vivo and this might serve as a molecular target for tumor therapy.

Background Dipeptidyl peptidase III (DPP3) is a zinc-dependent metallopeptidase and elevated in a variety of malignant tumors, but the underlying mechanism is not well understood so far. Here we investigated the association of esophageal carcinogenesis with the regulation of DPP3 expression by tissue-based quantitative analysis and the depletion of DPP3 expression in esophageal cancer cells and xenograft model. Methods The expression level of DPP3 in esophageal cancer tissues and adjacent normal tissues was detected in 93 cases of tissue biopsies collected from patients diagnosed with esophageal carcinoma by immunohistochemistry. The effect of DPP3 expression on cell proliferation, migration or apoptosis was determined in DPP3-depleted esophageal cancer cells created by infection with the lentivirus containing the shRNA specific to human DPP3 mRNA sequence followed by detection at the cellular level using celigo cell count assay, flow cytometry, wound-healing assay and trans-well assay as well as chip screening with a Human Apoptosis Antibody Array kit, which enables the quantitative detection of 43 apoptosis-related genes. A xenograft model was applied to the detection of tumor growth and invasion of DPP3-depleted cancer cells in nude mice. Results DPP3 expression was elevated in esophageal cancer tissues compared with adjacent non-tumor tissues (normal controls) with statistical significance (P < 0.05), and associated with poor prognosis of esophageal carcinoma. The DPP3-depletion resulted in a reduced cell proliferation and migration and enhanced cell-cycle arrest and apoptosis of esophageal cancer cells, and lead to the inhibition of tumor growth and invasion in xenograft model. In addition, DPP3-depletion was associated with the upregulation of pro-apoptotic proteins and the downregulation of anti-apoptotic proteins. Conclusions These findings suggest that DPP3 may promote cell proliferation, migration and survival of esophageal cancer cells in vitro, and tumor growth and invasion of esophageal carcinoma in vivo and this might serve as a molecular target for tumor therapy.

Background: Among women, cervical cancer (CC) is the fourth most prevalent cancer worldwide. Multiple bio-makers have been identified for tumor diagnosis and prognosis. However, no single bio-maker for CC detection exists that is both highly sensitive and specific. In this report, we explored the role of guanine nucleotide-binding protein subunit beta-2-like 1 (GNB2L1) in CC progression in Uyghur women. Methods: In the present study, 132 patients with cervical squamous cell carcinoma (CSCC), 56 cases with cervical intraepithelial neoplasia (CIN) were enrolled. GNB2L1 mRNA and protein expression levels were analyzed in CSCC, CIN, and normal cervical (NC) tissues and in CC cell lines using qRT-PCR, western blot, and immunohistochemistry. Stable GNB2L1-knockdown CC cells (shGNB2L1) were also created and lipid accumulation was measured with Oil Red O staining. Additionally, changes in GNB2L1 gene copy number were investigated with qRT-PCR and sequencing. A gene regulatory network was constructed using the STRING database. Finally, patient survival was assessed with Kaplan‑Meier and log-rank tests. Results: Our results indicated that GNB2L1 mRNA and protein are overexpressed in CSCC tumor tissues when compared to CIN and NC tissues. GNB2L1 expression was associated with the degree of cell differentiation (P = 0.001), lymph node metastasis (P = 0.049), and the clinical stage (P = 0.017). Oil Red O staining showed that lipid levels were lower in shGNB2L1 cells, and that GNB2L1 DNA copy number in CC tissues (29,7200 ± 9,815.061) was significantly higher when compared to NC tissues (19,5100 ± 5,676.392 [T=-2.770, P = 0.009]). Furthermore, patients with CSCC and high GNB2L1 expression demonstrated shorter overall survival times when compared to patients with low GNB2L1 expression (P = 0.002). Multivariate analysis indicated that GNB2L1 overexpression is an indepen­dent prognostic factor in patients with CSCC. Conclusions: Overexpression of GNB2L1 may contribute to tumor progression; therefore, it is a potential prognostic biomarker for CSCC patients.

Background Cervical cancer (CC), an aggressive squamous cell carcinoma, is characterized by early lymph node metastasis and extremely poor prognosis. We previously showed aberrant glycolysis in patients with CC. The upregulation of Receptor for Activated C Kinase 1 (RACK1) is associated with CC lymph node metastasis. However, its role in mediating aerobic glycolysis in CC lymph node metastasis remains unclear. Methods 104 cervical cancer and 31 cervicitis tissue were enrolled. RACK1, insulin-like growth factor 1 receptor (IGF1R), POU class 2 homeobox 2 (POU2F2) and Hexokinase 2 (HK2) expression in cervical cancer patients were detected by Immunohistochemistry analysis. 1H-NMR experiment was used for metabolomics of cell supernatant after RACK1 knockdown. Tube formation assay, Transwell assay, determination of glucose consumption and lactate production assay were performed to detected MS751 and SiHa cells lymphangiogenesis, migration, invasion, glucose consumption and lactate production. Chromatin immunoprecipitation-PCR assay and Dual-Luciferase Reporter Assay was performed to detect the binding among POU2F2 and RACK1. Co-immunoprecipitation and Immunofluorescence staining were performed to detect the binding among RACK1 and GF1R. Xenograft footpad lymph node metastasis was performed using nude mice. Xenograft tumor and metastatic lymph node tissues of mice were experienced immunohistochemistry and hematoxylin–eosin staining. Results Here, 1H-NMR analysis revealed significantly correlated RACK1 expression with glycolysis/gluconeogenesis pathway. Additionally, RACK1 knockdown inhibited aerobic glycolysis and lymphangiogenesis in vitro and suppressed CC lymph node metastasis in vivo. Furthermore, AKT/mTOR signaling was identified as a critical RACK1-regulated pathway that led to increased lymphangiogenesis in CC. Co-immunoprecipitation, immunofluorescence, and Western blot showed that RACK1 activated AKT/mTOR signaling by interacting with IGF1R. POU2F2 binds to RACK1 promoter and regulates RACK1 transcription, thus functionally contributing to glycolysis and lymphangiogenesis in CC. Importantly, administration of 2-deoxy-D-glucose, which attenuates glycolysis, inhibited RACK1-induced lymphangiogenesis in CC. The positive correlations between RACK1, IGF1R, POU2F2, and HK2 were further confirmed in CC tissues. Conclusion RACK1 plays an important role in CC tumor lymph node metastasis by regulating IGF1R/AKT/mTOR signaling mediated glycolysis pathways. Targeting the POU2F2/ RACK1/IGF1R/AKT/mTOR signaling pathway may provide a novel strategy for CC treatment.