Immune-related long noncoding RNA zinc finger protein 710-AS1-201 promotes the metastasis and invasion of gastric cancer cells.

BACKGROUND: Gastric cancer (GC) is a prevalent malignant tumor of the gastrointestinal system. ZNF710 is a transcription factor (TF), and zinc finger protein 710 (ZNF710)-AS1-201 is an immune-related long noncoding RNA (lncRNA) that is upregulated in GC cells. AIM: To assess the correlation between ZNF710-AS1-201 and immune microenvironment features and to investigate the roles of ZNF710-AS1-201 in the invasion and metastasis processes of GC cells. METHODS: We obtained data from The Cancer Genome Atlas and Wujin Hospital. We assessed cell growth, migration, invasion, and programmed cell death using cell counting kit-8, EdU, scratch, Transwell, and flow cytometry assays. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to identify the potential downstream targets of ZNF710-AS1-201. RESULTS: In GC tissues with low ZNF710-AS1-201 expression, immunoassays detected significant infiltration of various antitumor immune cells, such as memory CD8 T cells and activated CD4 T cells. In the low-expression group, the half-maximal inhibitory concentrations (IC50s) of 5-fluorouracil, cisplatin, gemcitabine, and trametinib were lower, whereas the IC50s of dasatinib and vorinostat were higher. The malignant degree of GC was higher and the stage was later in the high-expression group. Additionally, patients with high expression of ZNF710-AS1-201 had lower overall survival and disease-free survival rates. In vitro, the overexpression of ZNF710-AS1-201 greatly enhanced growth, metastasis, and infiltration while suppressing cell death in HGC-27 cells. In contrast, the reduced expression of ZNF710-AS1-201 greatly hindered cell growth, enhanced apoptosis, and suppressed the metastasis and invasion of MKN-45 cells. The expression changes in ZNF710 were significant, but the corresponding changes in isocitrate dehydrogenase-2, Semaphorin 4B, ARHGAP10, RGMB, hsa-miR-93-5p, and ZNF710-AS1-202 were not consistent or statistically significant after overexpression or knockdown of ZNF710-AS1-201, as determined by qRT-PCR. CONCLUSION: Immune-related lncRNA ZNF710-AS1-201 facilitates the metastasis and invasion of GC cells. It appears that ZNF710-AS1-201 and ZNF710 have potential as effective targets for therapeutic intervention in GC. Nevertheless, it is still necessary to determine the specific targets of the ZNF710 TF.

Crosstalk between mesenchymal stem cells and macrophages in inflammatory bowel disease and associated colorectal cancer.

Mesenchymal stem cells (MSCs) are attractive seed cells for immunotherapy, tissue engineering and regenerative medicine due to their self-renewal and multidirectional differentiation abilities, diverse immunoregulatory functions and ease of isolation from a wide range of tissues. MSCs exert their immunoregulatory effect on immune cells via cell-to-cell contact and paracrine mechanisms. In turn, MSCs can also be modulated by immune cells. Macrophages are constantly present in the mucosa of the intestinal tract of mammals and play an important role in the development and progression of inflammatory bowel disease (IBD), a chronic and recurrent inflammatory disease of the gastrointestinal tract characterized by idiopathic mucosal inflammation. The increased morbidity and mortality of IBD have made it a disease hard to cure in the clinic. MSCs have emerged as an important tool for IBD therapy due to their abilities to differentiate into enterocyte-like cells and regulate inflammatory cells, especially macrophages. In this review, we discuss the recent advances in the interaction between MSCs and macrophages in diseases, with an emphasis on IBD. We propose that an optimized MSC-based therapy would provide a novel strategy for the treatment of IBD and the prevention of IBD-associated colorectal cancer (CRC).

Enantioselective total synthesis of (+)-Lingzhiol via tandem semipinacol rearrangement/Friedel-Crafts type cyclization.

Enantioselective total synthesis of (+)-Lingzhiol has been achieved. It is the first example of in tandem semipinacol rearrangement reactions, the migrated aryl group further reacting with the carbonyl oxonium electrophile to furnish a polycyclic skeleton. Our synthesis involves 13 steps and proceeds in 6% overall yield.

Concise synthesis of (±)-Lingzhiol via epoxy-arene cyclization.

Concise synthesis of (±)-Lingzhiol has been achieved. The key reaction involves one-step construction of a 5/5/6/6 tetra-ring backbone of Lingzhiol via epoxy-arene cyclization.

Sesquiterpenoids with new carbon skeletons from the resin of Toxicodendron vernicifluum as new types of extracellular matrix inhibitors.

Toxicodenanes A-C (1-3), representing sesquiterpenoids with three new carbon skeletons, were isolated from the dried resin of Toxicodendron vernicifluum. Their structures were identified by spectroscopic data and X-ray diffraction crystallography. Their plausible biosynthetic route was proposed via the isolated intermediate (4). Compounds 2 and 3 could significantly inhibit overproduction of fibronectin, collagen IV, and IL-6 in high-glucose-induced mesangial cells in a dose- and time-dependent manner, showing their potential in diabetic nephropathy.

[Inhibition of culture supernatant of mesenchymal stem cells on macrophages RAW264.7 activated by soluble egg antigen of Schistosoma japonicum].

OBJECTIVE: To observe the inhibitive effect of rat mesenchymal stem cells (MSC) culture supernatant on macrophages activated by soluble egg antigen (SEA) of Schistosoma japonicum. METHODS: To select optimal SEA effecting concentration and time, macrophages RAW264.7 were induced by 5, 10, 20 or 40 microg/ml SEA for 12 h, or by 20 microg/ml SEA for 4, 8, 12 or 24 h before examination of TNF-alpha mRNA by RT-PCR. Macrophages were divided into five groups, i.e. negative control group, SEA group, SEA+MSC supernatant group (MSC group), SEA+NRK-52E supernatant group (NRK-52E group) and SEA+DMEM group (DMEM group). Except negative control group, macrophages in other four groups were induced by 20 microg/ml SEA for 12 h. SEA was then removed from MSC group, NRK-52E group and DMEM group and replaced with MSC supernatant, NRK-52E supernatant and DMEM, respectively. Morphology of macrophages in each group was observed by microscope after cultured with supernatant for 12 h. TNF-alpha mRNA in macrophages was detected by real-time quantitative PCR after cultured with supernatant for 12 h and 24 h. TGF-beta1 in macrophages was observed by Western blotting analysis after cultured with supernatant for 12 h. Macrophage proliferation was tested by MTT method after cultured with supernatant for 24 h and 48 h. RESULTS: The optimal SEA concentration and time for macrophage activation was 20 microg/ml and 12 h, respectively. Compared with SEA group, NRK-52E group, and DMEM group, macrophages in MSC group were round and small with less pseudopodia after cultured with supernatant for 12 h. TNF-alpha mRNA after cultured with MSC supernatant for 12 h and 24 h was (1.0 +/- 0.4) and (1.0 +/- 0.5) fold of negative control group, respectively, significantly less than NRK-52E group [(10.4 +/- 3.9) and (16.5 +/- 5.0) fold] (12 h: P < 0.05; 24 h: P < 0.01) and DMEM group [(6.0 +/- 2.1) and (2.4 +/- 0.7) fold] (P < 0.05). The grey density image analysis of TGF-beta1/GAPDH was 0.31 +/- 0.10 in MSC group, much lower than 0.88 +/- 0.10 in NRK-52E group (P < 0.01) and 0.58 +/- 0.06 in DMEM group (P < 0.05) after cultured with supernatant for 12 h. After 48 h culture, A490 of macrophages in MSC group was 0.22 +/- 0.05, much lower than 0.53 +/- 0.02 in NRK-52E group and 0.31 +/- 0.03 in DMEM group (P < 0.05). CONCLUSION: MSC supernatant can inhibit activation and proliferation of macrophages which were induced by SEA of S. japonicum.

Immunosuppressive effects of mesenchymal stem cells in collagen-induced mouse arthritis.

OBJECTIVE: The objective of this study was to investigate the efficacy of mesenchymal stem cell (MSC) in the treatment of arthritis. METHODS: Mesenchymal stem cells were injected intravenously into mice with collagen-induced arthritis (CIA). Arthritic indexes were evaluated, and the levels of the pro- and anti-inflammatory cytokines interleukin-10 (IL-10), gamma interferon (IFN-gamma)-inducible protein 10 (IP-10), chemokine (C-X-C motif) receptor 3 (CXCR3), interleukin-17A (IL-17A), and tumor necrosis factor alpha (TNF-alpha) in serum or splenic cells were determined using real-time RT-PCR, Western blotting, and enzyme-linked immunosorbent assay (ELISA). The proliferation of dendritic cell line D2SC cells was determined using (3)H-TdR incorporation assay. RESULTS: Upon injection of MSCs, overall arthritis symptoms were significantly improved in the CIA mouse models as indicated by the paw edema. Consistent with this observation, serum levels of pro-inflammatory cytokine TNF-alpha and inflammatory cell infiltration decreased significantly 12 days after MSC injection, while the expression of anti-inflammatory cytokines IL-10, IP-10, and CXCR3 was increased in splenocytes. In addition, we provided evidence that MSCs may directly promote the proliferation of D2SC cells and the expression of IP-10 in D2SC cells in vitro. CONCLUSION: Mesenchymal stem cells significantly enhance the efficacy of collagen-induced arthritis treatment, likely through the modulation of the expression of various cytokines.

[The expression of Toll-like receptor 8 and cytokines in rheumatoid arthritis mice induced by chicken II collogen].

AIM: To explore the expression of Toll-like receptor 8(TLR8)in rheumatoid arthritis induced by chicken II collogen in mice and analyze the relation of TLR8 to IL-10, IL-17A and IL-1beta. METHODS: Twelve DBA/1J mice were randomly divided into model group(T) and negative group(NC). The paw swelling was seen at the day of 27 post the first immunization with chicken II collogen. The serum TNF-alpha was detected by ELISA and inflammatory cell infiltration was examined by HE. Real-time PCR was used to analysis the expression of IL-10, IL-17A, IL-1beta and TLR8 in spleen mononuclear cells. RESULTS: Severe inflammation was detected in model group mice by histological analysis, which was not seen in negative group. The serum TNF-alpha in model group was enhanced compared with that in negative group(P<0.01). The expression levels of IL-10, IL-17A, TLR8 and IL-1beta in spleen mononuclear cells from model group were increased compared with those from negative group(P<0.01, P<0.05). Statistical analysis showed that there was negative correlation between the expression of TLR8 and IL-10(P<0.01), and there was no correlation between the expression of TLR8 and IL-17A or IL-1beta(P>0.05). CONCLUSION: The expression of TLR8 was increased in rheumatoid arthritis-mice induced by chicken II collogen, and there was negative correlation between the expression of TLR8 and the expression of IL-10.

Oct4, a novel marker for human gastric cancer.

BACKGROUND AND OBJECTIVE: Octamer-4 (Oct4), a transcription factor involved in regulating human embryonic stem cells (ESCs), may play a role in tumorigenesis. Since little is known about the efficacy of Oct4 as a potential biomarker for gastric cancer (GC), we investigated its expression in GC tissues and its relationship to various clinicopathological parameters. METHODS: Primary tumor tissues and matching, adjacent non-cancerous tissues were obtained from 62 GC patients, and Oct4 expression was examined by reverse transcription-PCR (RT-PCR) and real-time PCR. Twenty biopsy specimens of atrophic gastritis and gastric ulcer individually were collected as control. To detect Oct4 expression in the paired GC and non-cancerous tissues at the protein level, Western blotting and immunohistochemistry (IHC) were employed. Correlation analyses were conducted to assess the relationship between Oct4 expression and clinicopathological parameters. RESULTS: Oct4 expression levels were higher in GC tissues compared to matching, adjacent non-cancerous tissues, atrophic gastritis and gastric ulcer tissues. Additionally, Oct4 expression in GC tumors correlated with their differentiation status, but not with patient age or gender, tumor size, TNM stage, depth of invasion, or the presence of lymph node metastasis. CONCLUSIONS: Oct4 may be a potential biomarker for the initiation, progression, and differentiation of human GC.

[Expression of human phosphodiesterase 3A gene using baculovirus expression system in insect cell].

AIM: To investigate the expression of recombinant human phosphodiesterase 3A (HPDE3A) using baculovirus expression system in Tn cell line. METHODS: The HPDE3A cDNA was recombined with baculovirus, and then the recombinant was transfected into Tn cell line. The expression of HPDE3A in Tn cell line was detected and identified by the RT-PCR, SDS-PAGE and Western blotting. RESULTS: The recombinant HPDE3A protein was stably expressed in Tn cell line and detected by the distinct morphological changes of Tn cell, RT-PCR, SDS-PAGE and Western blotting using polyclonal antibody. The M(w) of the recombinant protein was about 120 kD. CONCLUSION: Recombinant HPDE3A can be expressed in Tn cell line using the baculovirus expression system, and thus provided the basic material for studying its bioactivity and application in screening for HPDE3A inhibitor.