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OBJECTIVE@#To explore the therapeutic mechanism of Liushen Wan (LSW) against colitis-associated colorectal cancer (CAC) by network pharmacology.@*METHODS@#TCMSP, BATMAN-TCM, CNKI, PubMed, Genecards, OMIM, and TTD databases were used to obtain the related targets of LSW and CAC. The common targets of LSW and CAC were obtained using Venny online website. The PPI network was constructed using Cytoscape 3.8.2 to screen the core targets of LSW in the treatment of CAC. GO and KEGG enrichment analysis were conducted using DAVID database. The therapeutic effect of LSW on CAC was evaluated in a C57BL/6J mouse model of AOM/DSS-induced CAC by observing the changes in body weight, disease activity index, colon length, and size and number of the tumor. HE staining and RT-qPCR were used to analyze the effect of LSW on inflammatory mediators. Immunohistochemistry and TUNEL staining were used to evaluate the effect of LSW on the proliferation and apoptosis of AOM/DSS-treated colon tumor cells. Immunohistochemistry and Western blotting were used to detect the effects of LSW on the expression of TLR4 proteins in CAC mice.@*RESULTS@#Network pharmacology analysis identified 69 common targets of LSW and CAC, and 33 hub targets were screened in the PPI network. KEGG pathway enrichment analysis suggested that the effect of LSW on CAC was mediated by the Toll-like receptor signaling pathway. In the mouse model of AOM/DSS-induced CAC, LSW significantly inhibited colitis-associated tumorigenesis, reduced tumor number and tumor load (P < 0.05), obviously improved histopathological changes in the colon, downregulated the mRNA levels of proinflammatory cytokines, and inhibited the proliferation (P < 0.01) and promoted apoptosis of colon tumor cells (P < 0.001). LSW also significantly decreased TLR4 protein expression in the colon tissue (P < 0.05).@*CONCLUSION@#LSW can inhibit CAC in mice possibly by regulating the expression of TLR4 to reduce intestinal inflammation, inhibit colon tumor cell proliferation and promote their apoptosis.
Subject(s)
Mice , Animals , Toll-Like Receptor 4 , Colitis-Associated Neoplasms , Network Pharmacology , Mice, Inbred C57BL , Colonic Neoplasms/pathologyABSTRACT
Aim To investigate the effects of long non-coding RNA(lncRNA)UNC5B-AS1 on the proliferation and epithelial mesenchymal transformation(EMT)of cervical cancer. Methods GEO and TCGA databases were used to download data sets and differential expression analysis was performed. qRT-PCR was used to verify the differential expression of lncRNA UNC5B-AS1 in normal and cancerous cervical tissues.The interference and overexpression of lncRNA UNC5B-AS1 were transfected into cervical cancer cell lines, and plate cloning, CCK-8 and EdU experiments were used to detect the effect of lncRNA UNC5B-AS1 on the pro-liferation of cervical cancer cells.Transwell assay was used to detect its effect on migration and invasion of cervical cancer cells.The expression levels of EMT-related genes E-Cadherin, N-Cadherin and Vimentin were detected by Western blot. Transcriptome sequencing was used to obtain the signal pathway regulated by lncRNA UNC5B-AS1, and to verify the expression level of related genes. Results RNA microarray and bioinformatics analysis showed that the expression level of lncRNA UNC5B-AS1 in cervical cancer was significantly higher than that in normal cervical tissue, and correlated with the overall survival time of patients.Compared with the negative control group, knockdown lncRNA UNC5B-AS1 could reduce the proliferation, migration and invasion of cervical cancer cells, while overexpression could promote the proliferation, migration and invasion of cervical cancer cells. Western blot showed that lncRNA UNC5B-AS1 could regulate EMT of cervical cancer cells. Transcriptome sequencing showed that lncRNA UNC5B-AS1 could regulate Toll like receptor(TLR)signaling pathway. qRT-PCR and Western blot results showed that the expression levels of TLR-related genes IL-6 and TICAM2 in the knockdown and overexpression lncRNA UNC5B-AS1 group were significantly changed(P<0.05). Conclusions LncRNA UNC5B-AS1 is highly expressed in cervical cancer. Overexpression of lncRNA UNC5B-AS1 may enhance TLR signaling pathway activity, thereby promoting proliferation and EMT of cervical cancer cells.
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Objective: To predict the targets and pathways for the main active components of Smilax glabra in the treatment of gout based on network pharmacology. Methods The active components and targets of the S. glabra were obtained by TCMSP database and Drugbank database. Furthemore, the interaction network among the targets was established by Cytoscape software. Meanwhile, crosslink analysis was performed to screen out the active components and potential targets. Finally, the information was obtained from the MAS 3.0 biomolecular function system, and then the target pathway network model was established. Results In this study, a total of 11 effective components and 39 effective targets were predicted, which related to adipocytokine signaling pathway, ERbB signaling pathway, and Toll like receptor signaling pathway. Among these pathways, MAPK1, RELA, PTGS2 genes may play a crucial role. Conclusion This study investigated the characteristics on multi-component, multi-target and multi-pathway of S. glabra, which provided a new idea and method for further study on anti-gout mechanism of S. glabra.
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Toll-like receptors(TLRs) signaling pathways are involved in the activation of innate and adaptive immune re-sponse and play an important role in the development of colorectal cancer. Deregulation of TLRs signaling pathway can lead to epitheli-al hemorrhage,chronic inflammation and the development of colorectal cancer. There is now lots of evidence that targeting this pathway will benefit the treatment of colorectal cancer,such as BCG,monophosphatidyl A,and iquimod,etc. ,which are now in clinical use. This article reviews current different function of TLRs in tumor development and their application in the treatment of colorectal cancer.
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Objective To identify a myeloid differentiation factor 88(MyD88)in Oncomelania hupensis,and characterize the role of MyD88 against Schistosoma japonicum infection. Methods The complete cDNA of MyD88 in O. hupensis was ob-tained by using rapid amplification of cDNA ends(RACE),and homologues sequences and conserved domains were aligned and the structure of MyD88 was predicted either. A phylogenetic tree of MyD88 was further constructed with other species. In ad-dition,the mRNA expression level of O. hupensis MyD88 before and after S. japonicum infection was investigated by real-time quantitative PCR(RT-qPCR). Results The cDNA of O. hupensis MyD88 consisted of 1406 bp open reading frame(ORF),en-coding 468 amino acid residues,which contained death domain and Toll/interlrukin-1 receptor(TIR)domain,the typical fea-tures of MyD88 family proteins. The predicted amino acid sequence of O. hupensis MyD88 shared 38%-52%identity with other mollusc. O. hupensis MyD88 was phylogenetically closeted to Biomphalaria glabrata MyD88. The O. hupensis MyD88 existed in all selected tissues and expressed highly in hemocyte,up-regulated after S. japonicum infection in all selected tissues except cephalopodium,especially higher in whole snail and hemocyte. Conclusion MyD88-dependent signaling pathway is present in O. hupensis and plays an important role in innate immune response against S. japonicum infection.