Background: DOK3 (Downstream of kinase 3) is involved primarily with immune cell infiltration. Recent research reported the role of DOK3 in tumor progression, with opposite effects in lung cancer and gliomas;however, ...Background: DOK3 (Downstream of kinase 3) is involved primarily with immune cell infiltration. Recent research reported the role of DOK3 in tumor progression, with opposite effects in lung cancer and gliomas;however, its role in prostate cancer (PCa) remains elusive. This study aimed to explore the role of DOK3 in PCa and to determine the mechanisms involved. Methods: To investigate the functions and mechanisms of DOK3 in PCa, we performed bioinformatic and biofunctional analyses. Samples from patients with PCa were collected from West China Hospital, and 46 were selected for the final correlation analysis. A lentivirus-based short hairpin ribonucleic acid (shRNA) carrier was established for silencing DOK3. A series of experiments involving the cell counting kit-8, bromodeoxyuridine, and flow cytometry assays were performed to identify cell proliferation and apoptosis. Changes in biomarkers from the nuclear factor kappa B (NF-κB) signaling pathway were detected to verify the relationship between DOK3 and the NF-κB pathway. A subcutaneous xenograft mouse model was performed to examine phenotypes after knocking down DOK3 in vivo . Rescue experiments with DOK3 knockdown and NF-κB pathway activation were designed to verify regulating effects. Results: DOK3 was up-regulated in PCa cell lines and tissues. In addition, a high level of DOK3 was predictive of higher pathological stages and worse prognoses. Similar results were observed with PCa patient samples. After silencing DOK3 in PCa cell lines 22RV1 and PC3, cell proliferation was significantly inhibited while apoptosis was promoted. Gene set enrichment analysis revealed that DOK3 function was enriched in the NF-κB pathway. Mechanism experiments determined that knockdown of DOK3 suppressed activation of the NF-κB pathway, increased the expressions of B-cell lymphoma-2 like 11 (BIM) and B-cell lymphoma-2 associated X (BAX), and decreased the expression of phosphorylated-P65 and X-linked inhibitor of apoptosis (XIAP). In the rescue experiments, pharmacological activation of NF-κB by tumor necrosis factor-α (TNF-α) partially recovered cell proliferation after the knockdown of DOK3. Conclusion: Our findings suggest that overexpression of DOK3 promotes PCa progression by activating the NF-κB signaling pathway.展开更多
Using suppression subtractive hybridization, a renal cell carcinoma (RCC) cDNA subtractive library which only contains differently expressed cDNAs between human RCC and normal kidney has been constructed. 200 clones w...Using suppression subtractive hybridization, a renal cell carcinoma (RCC) cDNA subtractive library which only contains differently expressed cDNAs between human RCC and normal kidney has been constructed. 200 clones were picked out randomly to perform enzyme digest analysis, a part of them underwent sequence analysis and Northern blot to identify RCC specially expressed genes. Results showed that 190 clones contain 50-400 bp inserts respectively. Sequence analysis was performed in 10 clones. All the 10 sequences were unknown before and derived from 6 unique novel genes among which the cDNA insert RCC18 has five copies. Northern blot analysis showed that RCC18 cDNA expressed highly in RCC, but there was no signal detected in normal kidney, and the full length of RCC18 was about 2.5 kb. The constructed cDNA subtractive library of human RCC is a highly efficient one and lays the solid foundation for large-scale screening and cloning new and specific oncogenes or tumor suppressor genes of RCC. The novel展开更多
Cellular mechanotransduction,a critical regulator of numerous biological processes,is the conversion from mechanical signals to biochemical signals regarding cell activities and metabolism.Typical mechanical cues in o...Cellular mechanotransduction,a critical regulator of numerous biological processes,is the conversion from mechanical signals to biochemical signals regarding cell activities and metabolism.Typical mechanical cues in organisms include hydrostatic pressure,fluid shear stress,tensile force,extracellular matrix stiffness or tissue elasticity,and extracellular fluid viscosity.Mechanotransduction has been expected to trigger multiple biological processes,such as embryonic development,tissue repair and regeneration.However,prolonged excessive mechanical stimulation can result in pathological processes,such as multi-organ fibrosis,tumorigenesis,and cancer immunotherapy resistance.Although the associations between mechanical cues and normal tissue homeostasis or diseases have been identified,the regulatory mechanisms among different mechanical cues are not yet comprehensively illustrated,and no effective therapies are currently available targeting mechanical cue-related signaling.This review systematically summarizes the characteristics and regulatory mechanisms of typical mechanical cues in normal conditions and diseases with the updated evidence.The key effectors responding to mechanical stimulations are listed,such as Piezo channels,integrins,Yes-associated protein(YAP)/transcriptional coactivator with PDZ-binding motif(TAZ),and transient receptor potential vanilloid 4(TRPV4).We also reviewed the key signaling pathways,therapeutic targets and cutting-edge clinical applications of diseases related to mechanical cues.展开更多
This study aimed to further validate the prognostic role of fibrinogen in upper tract urothelial carcinoma(UTUC)in a large Chinese cohort.A total of 703 patients who underwent radical nephroureterectomy were retrospec...This study aimed to further validate the prognostic role of fibrinogen in upper tract urothelial carcinoma(UTUC)in a large Chinese cohort.A total of 703 patients who underwent radical nephroureterectomy were retrospectively identified.Fibrinogen levels of≥4.025 g l?1 were defined as elevated.Logistic regression analysis was performed to determine the association between fibrinogen and adverse pathological features.Kaplan–Meier analysis and Cox regression models were used to assess the associations of fibrinogen with cancer-specific survival(CSS),disease recurrence-free survival(RFS),and overall survival(OS).Harrell c-index and decision curve analysis were used to assess the clinical utility of multivariate models.The median follow-up duration was 42(range:1–168)months.Logistic regression analysis revealed that elevated fibrinogen was associated with higher tumor stage and grade,lymph node involvement,lymphovascular invasion,sessile carcinoma,concomitant variant histology,and positive surgical margins(all P<0.05).Multivariate Cox regression analysis demonstrated that elevated fibrinogen was independently associated with decreased CSS(hazard ratio[HR]:2.33;P<0.001),RFS(HR:2.09;P<0.001),and OS(HR:2.09;P<0.001).The predictive accuracies of the multivariate models were improved by 3.2%,2.0%,and 2.8%for CSS,RFS,and OS,respectively,when fibrinogen was added.Decision curve analysis showed an added benefit for CSS prediction when fibrinogen was added to the model.Preoperative fibrinogen may be a strong independent predictor of worse oncologic outcomes in UTUC;therefore,it may be valuable to apply this marker to the current risk stratification in UTUC.展开更多
基金supported by the National Key Research and Development Program of China(No.2017YFC0908003).
文摘Background: DOK3 (Downstream of kinase 3) is involved primarily with immune cell infiltration. Recent research reported the role of DOK3 in tumor progression, with opposite effects in lung cancer and gliomas;however, its role in prostate cancer (PCa) remains elusive. This study aimed to explore the role of DOK3 in PCa and to determine the mechanisms involved. Methods: To investigate the functions and mechanisms of DOK3 in PCa, we performed bioinformatic and biofunctional analyses. Samples from patients with PCa were collected from West China Hospital, and 46 were selected for the final correlation analysis. A lentivirus-based short hairpin ribonucleic acid (shRNA) carrier was established for silencing DOK3. A series of experiments involving the cell counting kit-8, bromodeoxyuridine, and flow cytometry assays were performed to identify cell proliferation and apoptosis. Changes in biomarkers from the nuclear factor kappa B (NF-κB) signaling pathway were detected to verify the relationship between DOK3 and the NF-κB pathway. A subcutaneous xenograft mouse model was performed to examine phenotypes after knocking down DOK3 in vivo . Rescue experiments with DOK3 knockdown and NF-κB pathway activation were designed to verify regulating effects. Results: DOK3 was up-regulated in PCa cell lines and tissues. In addition, a high level of DOK3 was predictive of higher pathological stages and worse prognoses. Similar results were observed with PCa patient samples. After silencing DOK3 in PCa cell lines 22RV1 and PC3, cell proliferation was significantly inhibited while apoptosis was promoted. Gene set enrichment analysis revealed that DOK3 function was enriched in the NF-κB pathway. Mechanism experiments determined that knockdown of DOK3 suppressed activation of the NF-κB pathway, increased the expressions of B-cell lymphoma-2 like 11 (BIM) and B-cell lymphoma-2 associated X (BAX), and decreased the expression of phosphorylated-P65 and X-linked inhibitor of apoptosis (XIAP). In the rescue experiments, pharmacological activation of NF-κB by tumor necrosis factor-α (TNF-α) partially recovered cell proliferation after the knockdown of DOK3. Conclusion: Our findings suggest that overexpression of DOK3 promotes PCa progression by activating the NF-κB signaling pathway.
基金This work was supported by the National Natural Science Foundation of China (Grant No. 39870841).
文摘Using suppression subtractive hybridization, a renal cell carcinoma (RCC) cDNA subtractive library which only contains differently expressed cDNAs between human RCC and normal kidney has been constructed. 200 clones were picked out randomly to perform enzyme digest analysis, a part of them underwent sequence analysis and Northern blot to identify RCC specially expressed genes. Results showed that 190 clones contain 50-400 bp inserts respectively. Sequence analysis was performed in 10 clones. All the 10 sequences were unknown before and derived from 6 unique novel genes among which the cDNA insert RCC18 has five copies. Northern blot analysis showed that RCC18 cDNA expressed highly in RCC, but there was no signal detected in normal kidney, and the full length of RCC18 was about 2.5 kb. The constructed cDNA subtractive library of human RCC is a highly efficient one and lays the solid foundation for large-scale screening and cloning new and specific oncogenes or tumor suppressor genes of RCC. The novel
基金This work was supported by the National Science Fund of China(Grant Number:82270720)National Clinical Research Center for Geriatrics,West China Hospital,Sichuan University(Grant Number:Y2021LC005)+1 种基金1.3.5 project for disciplines of excellence,West China Hospital,Sichuan University(Grant Number:ZYGD18011)The National Science Fund of China(Grant Number:81873601).Many thanks to ClinicalTrials.gov(https://clinicaltrials.gov/ct2/home),which is maintained by the National Library of Medicine(NLM)at the National Institutes of Health(NIH),for the support of clinical trials collection.We appreciate the support from Biorender(https://www.biorender.com)for helping us create some of the figures.We thank Xintong Bao for her kind and excellent help in this study。
文摘Cellular mechanotransduction,a critical regulator of numerous biological processes,is the conversion from mechanical signals to biochemical signals regarding cell activities and metabolism.Typical mechanical cues in organisms include hydrostatic pressure,fluid shear stress,tensile force,extracellular matrix stiffness or tissue elasticity,and extracellular fluid viscosity.Mechanotransduction has been expected to trigger multiple biological processes,such as embryonic development,tissue repair and regeneration.However,prolonged excessive mechanical stimulation can result in pathological processes,such as multi-organ fibrosis,tumorigenesis,and cancer immunotherapy resistance.Although the associations between mechanical cues and normal tissue homeostasis or diseases have been identified,the regulatory mechanisms among different mechanical cues are not yet comprehensively illustrated,and no effective therapies are currently available targeting mechanical cue-related signaling.This review systematically summarizes the characteristics and regulatory mechanisms of typical mechanical cues in normal conditions and diseases with the updated evidence.The key effectors responding to mechanical stimulations are listed,such as Piezo channels,integrins,Yes-associated protein(YAP)/transcriptional coactivator with PDZ-binding motif(TAZ),and transient receptor potential vanilloid 4(TRPV4).We also reviewed the key signaling pathways,therapeutic targets and cutting-edge clinical applications of diseases related to mechanical cues.
基金This program was supported by the National key Research and Development program of China(Grant No.SQ2017YFSF090096)the Prostate Cancer Foundation Young Investigator Award 2013,the National Natural Science Foundation of China(Grant No.81300627,No.81370855,No.81702536,and No.81770756)+1 种基金Programs from Science and Technology Department of Sichuan Province(Grant No.2018JY0089 and No.2017HH0063)Young Investigator Award of Sichuan University 2017(Grant No.2017SCU04A17).The funders had no role in patient selection,data extraction,statistical analysis or interpretation,writing of this article,or the decision to publish.
文摘This study aimed to further validate the prognostic role of fibrinogen in upper tract urothelial carcinoma(UTUC)in a large Chinese cohort.A total of 703 patients who underwent radical nephroureterectomy were retrospectively identified.Fibrinogen levels of≥4.025 g l?1 were defined as elevated.Logistic regression analysis was performed to determine the association between fibrinogen and adverse pathological features.Kaplan–Meier analysis and Cox regression models were used to assess the associations of fibrinogen with cancer-specific survival(CSS),disease recurrence-free survival(RFS),and overall survival(OS).Harrell c-index and decision curve analysis were used to assess the clinical utility of multivariate models.The median follow-up duration was 42(range:1–168)months.Logistic regression analysis revealed that elevated fibrinogen was associated with higher tumor stage and grade,lymph node involvement,lymphovascular invasion,sessile carcinoma,concomitant variant histology,and positive surgical margins(all P<0.05).Multivariate Cox regression analysis demonstrated that elevated fibrinogen was independently associated with decreased CSS(hazard ratio[HR]:2.33;P<0.001),RFS(HR:2.09;P<0.001),and OS(HR:2.09;P<0.001).The predictive accuracies of the multivariate models were improved by 3.2%,2.0%,and 2.8%for CSS,RFS,and OS,respectively,when fibrinogen was added.Decision curve analysis showed an added benefit for CSS prediction when fibrinogen was added to the model.Preoperative fibrinogen may be a strong independent predictor of worse oncologic outcomes in UTUC;therefore,it may be valuable to apply this marker to the current risk stratification in UTUC.
