Secreted Frizzled-Related Protein 5 Mediates Wnt5a Expression in Microcystin-Leucine-Arginine-Induced Liver Lipid Metabolism Disorder in Mice

Objective Microcystin-leucine-arginine(MC-LR)exposure induces lipid metabolism disorders in the liver.Secreted frizzled-related protein 5(SFRP5)is a natural antagonist of winglesstype MMTV integration site family,member 5A(Wnt5a)and an anti-inflammatory adipocytokine.In this study,we aimed to investigate whether MC-LR can induce lipid metabolism disorders in hepatocytes and whether SFRP5,which has anti-inflammatory effects,can alleviate the effects of hepatic lipid metabolism by inhibiting the Wnt5a/Jun N-terminal kinase(JNK)pathway.Methods We exposed mice to MC-LR in vivo to induce liver lipid metabolism disorders.Subsequently,mouse hepatocytes that overexpressed SFRP5 or did not express SFRP5 were exposed to MC-LR,and the effects of SFRP5 overexpression on inflammation and Wnt5a/JNK activation by MC-LR were observed.Results MC-LR exposure induced liver lipid metabolism disorders in mice and significantly decreased SFRP5 mRNA and protein levels in a concentration-dependent manner.SFRP5 overexpression in AML12cells suppressed MC-LR-induced inflammation.Overexpression of SFRP5 also inhibited Wnt5a and phosphorylation of JNK.Conclusion MC-LR can induce lipid metabolism disorders in mice,and SFRP5 can attenuate lipid metabolism disorders in the mouse liver by inhibiting Wnt5a/JNK signaling.

Hypermethylation and expression regulation of secreted frizzled-related protein genes in colorectal tumor

AIM： To investigate the functions of promoter hypermethylation of secreted frizzled-related proteins （sFRPs） genes in colorectal tumorigenesis and progression. METHODS： The promoter hypermethylation and expression of sFRP genes in 72 sporadic colorectal carcinomas, 33 adenomas, 18 aberrant crypt foci （ACF） and colorectal cancer cell lines RKO, HCT116 and SW480 were detected by methylation-specific PCR and reverse transcription PCR, respectively. RESULTS： None of the normal colorectal mucosa tissues showed methylated bands of any of four sFRP genes, sFRP1, 2, 4 and 5 were frequently methylated in colorectal carcinoma, adenoma and ACF （sFRP1 〉 85%, sFRP2 〉75%, sFRP5 〉 50%）, and the differences between three colorectal tissues were not significant （P 〉 0.05）. IVlethylation in colorectal tumors was more frequent than in normal mucosa and adjacent normal mucosa. The mRNA of sFRP1-5 genes was expressed in all normal colorectal mucosa samples. Expression of sFRP1, 2, 4 and 5 and sFRP1, 2 and 5 was downregulated in carcinoma and adenoma, respectively. The downregulation of sFRP2, 4 and 5 was more frequent in carcinoma than in adenoma. Expression of sFRP3 which promoter has no CpG island was downregulated in only a few of colorectal tumor samples （7/105）. The downregulation ofsFRP1, 2, 4 and 5 expression was significantly associated with promoter hypermethylation in colorectal tumor. After cells were treated by DAC/TSA combination, the silenced sFRP mRNA expression could be effectively re-expressed in colorectal cancer cell lines. CONCLUSION： Hypermethylation of sFRP genes is a common early event in the evolution of colorectal tumor, occurring frequently in ACF, which is regarded as the earliest lesion of multistage colorectal carcinogenesis. It appears to functionally silence sFRP genes expression. Methylation of sFRP1, 2 and 5 genes might serve as indicators for colorectal tumor.

Hypermethylation and aberrant expression of Wnt antagonist secreted frizzled-related protein 1 in gastric cancer

AIM： To identify the methylation of secreted frizzled-related protein 1 （SFRP1） in gastric cancer and to investigate the aberrant expression of SFRP1 and its correlation with the clinical pathological features of patients. METHODS： We determined SFRP1 methylation and SFRP1 mRNA expression in 3 gastric cancer cell lines SGC-7901, BGC-823, HGC-27, from 52 primary gastric cancer specimens and matched tumor adjacent tissue specimens by methylation-specific （MSP） PCR and RT-PCR respectively. Fisher＇s exact test was used to analyze the statistical association between clinical pathological data and aberrant expression of SFRP1. RESULTS： In 3 cancer cell lines, BGC-823 and HGC-27 had methylated SFRP1 and lost SFRP1 mRNA expression. After treatment of BGC-823 and HGC-27 with the demethylating agent, 5-aza-2′-deoxycytidine, SFRP1 was re-expressed. In 52 primary gastric cancer specimens and matched tumor adjacent tissue specimens, hypermethylation of SFRP1 was detected in 23 （44%） and 8 （15%） specimens respectively （x^2= 10.34, P 〈 0.01）. Loss of SFRP1 expression was detected in 17（33%） and 6 （12%） specimens respectively （x^2= 6.75, P 〈 0.01）. There was a significant correlation between SFRP1 hypermethylation and SFRP1 expression loss. SFRP1 expression was also correlated significantly with tumor stage and lymph node status, but not with patient sex, age and histological type. CONCLUSION： SFRP1 inactivation is a common and early event caused mainly by hypermethylation in gastric cancer. SFRP1 expression loss may be correlated with tumor metastasis in primary gastric cancer.

Secreted Frizzled-Related Protein 5 (SFRP5) in Patients with Obstructive Sleep Apnea

Objective Obstructive sleep apnea (OSA) is closely related to obesity, insulin resistance and inflammation. Secreted frizzled-related protein 5 (SFRP5) is a recently discovered adipokine. It is involved in insulin resistance and inflammation in obesity. This study aimed at evaluating the association between SFRP5and sleeping characteristics as well as biochemical parameters of OSA patients.Methods This was a prospective case control study. Nondiabetic OSA patients and controls were consecutively recruited and divided into three groups: OSA group, apnea–hypopnea Index (AHI)≥5/h; healthy controls with normal body mass index (BMI); obese controls without OSA, and BMI > 24.0 kg/m2. All participants underwent polysomnography (PSG). Plasma SFRP5 was examined using enzyme-linked immunosorbent assay (ELISA). Blood biochemical examinations, including fasting blood glucose (FBG), lipid profile, hypersensitive Creactive protein (hsCRP), were performed early in the morning after PSG. Patients with severe OSA were treated with nasal continuous positive airway pressure (nCPAP), and plasma SFRP5 was repeatedly measured for comparison.Results Sixty-eight subjects were enrolled in the study, including 38 patients of OSA, whose medium AHI was 58.70 /h (36.63, 71.15), 20 obese controls, and 10 healthy controls. The plasma SFRP5 level of OSA patients was not significantly different from that of healthy controls or obese controls. In OSA patients, SFRP5 level correlated positively with triglyceride level (r=0.447, P=0.005) and negatively with LDL-cholesterol level and HDLcholesterol level (r=?0.472 and P=0.003; r=?0.478 and P=0.002; respectively). SFRP5 level was not found correlating with FBG, AHI, or any of nocturnal hypoxia parameters. After overnight nCPAP treatment, plasma SFRP5 levels of OSA patients did not change significantly (t=1.557, P = 0.148) compared to that of pretreatment.Conclusions In nondiabetic OSA patients, plasma SFRP5 is associated with the lipid profile. However,no correlation was observed between SFRP5 and FBG or sleep parameters. The SFRP5 level of OSA patients did not differ from that of non-OSA individuals in our study.

Epigenetic inactivation of secreted frizzled-related protein 2 in esophageal squamous cell carcinoma

AIM： To investigate the expression and methylation status of the secreted frizzled-related protein 2 （SFRP2） in esophageal squamous cell carcinoma （ESCC） and ex- plore its role in ESCC carcinogenesis.METHODS： Seven ESCC cell lines （KYSE 30, KYSE150, KYSE410, KYSE510, EC109, EC9706 and TE-1） and one immortalized human esophageal epithelial cell line （Het- 1A）, 20 ESCC tissue samples and 20 paired adjacent non-tumor esophageal epithelial tissues were analyzed in this study. Reverse-transcription polymerase chain reaction （RT-PCR） was employed to investigate the expression of SFRP2 in cell lines, primary ESCC tumor tissue, and paired adjacent normal tissue. Methylation status was evaluated by methylation-specific PCR and bisulfite sequencing. The correlation between expres- sion and promoter methylation of the SFRP2 gene was confirmed with treatment of 5-aza-2＇-deoxycytidine. To assess the potential role of SFRP2 in ESCC, we es-tablished stable SFRP2-transfected cells and examined them with regard to cell proliferation, colony formation, apoptosis and cell cycle in vivo and in vitro.RESULTS： SFRP2 mRNA was expressed in the im- mortalized normal esophageal epithelial cell line but not in seven ESCC cell lines. By methylation-specific PCR, complete methylation was detected in three cell lines with silenced SFRP2 expression, and extensive methylation was observed in the other four ESCC cell lines. 5-aza-2＇-deoxycytidine could restore the expres- sion of SFRP2 mRNA in the three ESCC cell lines lack- ing SFRP2 expression. SFRP2 mRNA expression was obviously lower in primary ESCC tissue than in adjacent normal tissue （0.939 ± 0.398 vs 1.51 ± 0.399, P 〈 0.01）. SFRP2 methylation was higher in tumor tissue than in paired normal tissue （95% vs 65%, P 〈 0.05）. The DNA methylation status of the SFRP2 correlated inversely with the SFRP2 expression. To assess the potential role of SFRP2 in ESCC, we established stable SFRP2 transfectants and control counterparts by in- troducing pcDNA3.1/v5 hisA -SFRP2 or pcDNA3.1/v5 hisA -empty vector into KYSE30 cells lacking SFRP2 expression. After transfection, the forced-expression of SFRP2 was confirmed by the RT-PCR. In comparison with the control groups, stably-expressed SFRP2 in KYSE 30 cells significantly reduced colony formation in vitro （47.17% 4± 15.61% vs 17% ：1： 3.6%, P = 0.031） and tumor growth in nude mice （917.86：1：249.35 mm3 vs 337.23 ± 124.43 mm3, P 〈 0.05）. Using flow cytom- etry analysis, we found a significantly higher number of early apoptotic cells in SFRP2-transfected cells than in the control cells （P = 0.025）. The mean cell number in the S and G2-M phases of the cell cycle was also significantly lower in SFRP2-transfected KYSE30 cells compared with mock transfected counterparts. CONCLUSION： Silencing of SFRP2 expression through promoter hypermethylation may be a factor in ESCC carcinogenesis through loss of its tumor-suppressive activity.

Induction of CXC chemokines in human mesenchymal stem cells by stimulation with secreted frizzled-related proteins through non-canonical Wnt signaling

AIM: To investigate the effect of secreted frizzledrelated proteins(s FRPs) on CXC chemokine expression in human mesenchymal stem cells(h MSCs).METHODS: CXC chemokines such as CXCL5 and CXCL8 are induced in h MSCs during differentiation with osteogenic differentiation medium(OGM) and may be involved in angiogenic stimulation during bone repair. h MSCs were treated with conditioned medium(CM) from L-cells expressing non-canonical Wnt5 a protein, or with control CM from wild type L-cells, or directly with s FRPs for up to 10 d in culture. m RNA expression levels of both CXCL5 and CXCL8 were quantitated by real-time reverse transcriptase-polymerase chain reaction and secreted protein levels of these proteins determined by ELISA. Dose-(0-500 ng/m L) and time-response curves were generated for treatment with s FRP1. Signal transduction pathways were explored by western blot analysis with pan- or phosphorylation-specific antibodies, through use of specific pathway inhibitors, and through use of si RNAs targeting specific frizzled receptors(Fzd)-2 and 5 or thereceptor tyrosine kinase-like orphan receptor-2(Ro R2) prior to treatment with s FRPs. RESULTS: CM from L-cells expressing Wnt5 a, a noncanonical Wnt, stimulated an increase in CXCL5 m RNA expression and protein secretion in comparison to control L-cell CM. s FRP1, which should inhibit both canonical and non-canonical Wnt signaling, surprisingly enhanced the expression of CXCL5 at 7 and 10 d. Dickkopf1, an inhibitor of canonical Wnt signaling prevented the s FRPstimulated induction of CXCL5 and actually inhibited basal levels of CXCL5 expression at 7 but not at 10 d post treatment. In addition, all four s FRPs isoforms induced CXCL8 expression in a dose- and time-dependent manner with maximum expression at 7 d with treatment at 150 ng/m L. The largest increases in CXCL5 expression were seen from stimulation with s FRP1 or s FRP2. Analysis of mitogen-activated protein kinase signaling pathways in the presence of OGM showed s FRP1-induced phosphorylation of extracellular signal-regulated kinase(ERK)(p44/42) maximally at 5 min after s FRP1 addition, earlier than that found in OGM alone. Addition of a phospholipase C(PLC) inhibitor also prevented s FRPstimulated increases in CXCL8 m RNA. si RNA technology targeting the Fzd-2 and 5 and the non-canonical Fzd co-receptor Ro R2 also significantly decreased s FRP1/2-stimulated CXCL8 m RNA levels.CONCLUSION: CXC chemokine expression in h MSCs is controlled in part by s FRPs signaling through noncanonical Wnt involving Fzd2/5 and the ERK and PLC pathways.

Study on the Correlation Between SFRP-5 Expression Level, Insulin Resistance, and Glycolipid Metabolism in Gestational Diabetes Mellitus

Objective:To investigate the correlation between serum secretory frizzled-related protein 5(SFRP-5)expression levels and insulin resistance and glucolipid metabolism in patients with gestational diabetes mellitus(GDM).Methods:Baseline data were collected from 58 patients with GDM and 51 healthy controls who were admitted Affiliated Hospital of Hebei University from May 2020 to June 2022.sSTRA5 concentrations in peripheral blood of pregnant women were measured,and SFRP-5 levels in patients with different GDM types and normal controls were analyzed by logistic regression models.Results:The levels of triglyceride(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),fasting blood glucose(FBG),fasting insulin(FINS),hemoglobin A1c(HbA1c),and homeostasis model assessment-estimated insulin resistance(HOMA-IR)were higher in the observation group than in the control group,with statistically significant differences(P<0.05),while the expression levels of high-density lipoprotein cholesterol(HDL-C)and serum SFRP-5 were lower than in the control group,with statistically significant differences(P<0.05);serum SFRP-5,TG,TC,FBG,and HOMA-IR were all risk factors for GDM(P<0.05).Conclusion:Elevated serum sSTRA5 may be involved in the regulation of insulin resistance in the body and the regulation of blood glucose in the body by affecting lipid metabolism and inflammatory response.

Combination analysis of hypermethylated SFRP1 and SFRP2 gene in fecal as a novel epigenetic biomarker panel for colorectal cancer screening

Objective：To investigate the feasibility of the combination of detecting hypermethylated secreted frizzled-related protein 1 （SFRP1） and secreted frizzled-related protein 2（SFRP2） in feces as a panel of biomarkers for colorectal cancer（CRC） screening. Methods： Methylation-specific PCR（MSP） was performed to analyze methylation status of SFRP1 and SFRP2 in a blinded fashion in tumor tissues and in matched stool samples from 39 patients with primary CRC, 34 patients with adenomas, 17 patients with hyperplastic polyps and 20 endoscopically normal subjects as normal controls. Simultaneously we analyzed the correlation of hypermethylated SFRP1 and SFRP2 with the clinicopathological features of CRC. Results：Hypermethylated SFRP1 was detected in 92.3%, 76.5%, 47.1% of tissue samples and in 89.7%, 64.7%, 35.3% of matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. Hypermethylated SFRP2 was detected in 87.2%, 67.6%, 35.3% of tissue samples and in 82.1%, 55.9%, 29.4% of matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. Of these two genes, at least one hypermethylated was 94.9%, 82.4%, 52.9% in tissue samples and 92.3%,73.5%, 47.1% in matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. In contrast, no hypermethylated SFRP1 and SFRP2 were detected in mucosa tissues of normal controls, only 2 cases of fecal samples was detected with hypermethylated SFRP1 and another 1 case was detected with hypermethylated SFRP2. Moreover, no significant associations were observed between hypermethylated SFRP1,SFRP2 and clinicopathological features of CRC. Conclusion： Hypermethylation of SFRP1 and SFRP2 in feces are novel epigenetic biomarkers of CRC and carded high potential for the remote detection of CRC as non-invasive screening method, and combined analysis of hypermethylated SFRP1 and SFRP2 in fecal could further increase the detection rate of CRC and premalignant lesions.

肝细胞癌和慢性乙型肝炎患者血清sFRP-1和β-catenin水平及临床意义

目的探讨肝细胞癌(hepatocellular carcinoma,HCC)和慢性乙型肝炎(chronic hepatitis B,CHB)患者血清分泌型卷曲相关蛋白-1(secreted frizzled-related proteins,sFRP-1)和β-catenin水平及临床意义。方法采用ELISA法检测36例HCC、36例CHB和36例正常人血清sFRP-1和β-catenin水平,应用Spearman相关分析,分析血清sFRP-1和β-catenin水平与年龄、丙氨酸氨基转移酶(ALT)、白蛋白和总胆红素等的相关性。结果 HCC和CHB患者血清sFRP-1水平分别为(1136.28±332.43)pg/ml和(1194.53±156.84)pg/ml,均显著低于正常人[(1477.26±563.12)pg/ml,P<0.05];HCC患者血清β-catenin水平为(527.2±20.9)pg/ml,显著高于正常人和CHB患者[分别为(369.8±21.5)pg/ml和(374.0±8.3)pg/ml,P<0.05];HCC和CHB患者血清sFRP-1和β-catenin水平与年龄、ALT、白蛋白、总胆红素无显著相关性。结论肝细胞癌和慢性乙型肝炎患者血清sFRP-1降低,而肝细胞癌患者血清β-catenin水平升高。

Wnt signaling control of bone cell apoptosis

Wnts are a large family of growth factors that mediate essential biological processes like embryogenesis, morpho- genesis and organogenesis. These proteins also play a role in oncogenesis, and they regulate apoptosis in many tissues. Wnts bind to a membrane receptor complex comprised of a frizzled （FZD） G-protein-coupled receptor and a low-density lipoprotein （LDL） receptor-related protein （LRP）. The formation of this ligand-receptor complex initiates a number of signaling cascades that include the canonical/beta-catenin pathway as well as several noncanonical pathways. In recent years, canonical Wnt signaling has been reported to play a significant role in the control of bone formation. Clinical studies have found that mutations in LRP-5 are associated with reduced bone mineral density （BMD） and fractures. Investigations of knockout and transgenic mouse models of Wnt pathway components have shown that canonical Wnt signaling modulates most aspects ofosteoblast physiology including proliferation, differentiation, function and apoptosis. Transgenic mice expressing a gain of function mutant of LRP-5 in bone, or mice lacking the Wnt antagonist secreted frizzled-related protein-l, exhibit elevated BMD and suppressed osteoblast apoptosis. In addition, preclinical studies with pharmacologic compounds such as those that inhibit glycogen synthase kinase-3β support the importance of the canonical Wnt pathway in modulation of bone formation and osteoblast apoptosis.

Detection of aberrant methylation in fecal DNA as a molecular screening tool for colorectal cancer and precancerous lesions

AIM： To investigate the feasibility of detecting methylated fecal DNA as a screening tool for colorectal carcinoma （CRC） and precancerous lesions. METHODS： Methylated secreted frizzled-related protein gene 2 （SFRP2）, hyperplastic polyposis protein gene （HPP1） and O6-methylguanine-DNA methyltransferase gene （MGMT） in stools from 52 patients with CRC, 35 patients with benign colorectal diseases and 24 normal individuals were analyzed using methylation-specific PCR. RESULTS： Methylated SFRP2, HPP1 and MGMT were detected in 94.2%, 71.2%, 48.1% of CRC patients and 52.4%, 57.1%, 28.6% of adenoma patients, respectively. The overall prevalence of fecal DNA with at least one methylated gene was 96.2% and 81.8% in patients with CRC and precancerous lesions, respectively. In contrast, only one of the 24 normal individuals revealed methylated DNA. These results indicated a 93.7% sensitivity and a 77.1% specificity of the assay for detecting CRC and precancerous lesions. CONCLUSION： IVlethylation testing of fecal DNA using a panel of epigenetic markers may be a simple and promising non-invasive screening method for CRC and precancerous lesions.

Detection of promoter hypermethylation of Wnt antagonist genes in fecal samples for diagnosis of early colorectal cancer

AIM: To investigate the feasibility of detecting aberrantly hypermethylated Wnt-antagonist gene promoters (SFRP2 and WIF-1) in fecal DNA as non-invasive biomarkers for early colorectal cancer (CRC).

SFRP4 expression correlates with epithelial mesenchymal transitionlinked genes and poor overall survival in colon cancer patients

BACKGROUND Colon cancer is among the most commonly diagnosed cancers in the United States with an estimated 97220 new cases expected by the end of 2018.It affects 1.2 million people around the world and is responsible for about 0.6 million deaths every year.Despite decline in overall incidence and mortality over the past 30 years,there continues to be an alarming rise in early-onset colon cancer cases(<50 years).Patients are often diagnosed at late stages of the disease and tend to have poor survival.We previously showed that the WNT“gatekeeper”gene,secreted frizzled-related protein 4(SFRP4),is over-expressed in early-onset colon cancer.SFRP4 is speculated to play an essential role in cancer by inhibiting the epithelial mesenchymal transition(EMT).AIM To investigate the correlation between SFRP4 expression and EMT-linked genes in colon cancer and how it affects patient survival.METHODS SFRP4 expression relative to that of EMT-linked genes and survival analysis were performed using the University of California Santa Cruz Cancer Browser interface.RESULTS SFRP4 was found to be co-expressed with the EMT-linked markers CDH2,FN1,VIM,TWIST1,TWIST2,SNAI1,SNAI2,ZEB1,ZEB2,POSTN,MMP2,MMP7,MMP9,and COL1A1.SFRP4 expression negatively correlated with the EMTlinked suppressors CLDN4,CLDN7,TJP3,MUC1,and CDH1.The expression of SFRP4 and the EMT-linked markers was higher in mesenchymal-like samples compared to epithelial-like samples which potentially implicates SFRP4-EMT mechanism in colon cancer.Additionally,patients overexpressing SFRP4 presented with poor overall survival(P=0.0293).CONCLUSION Considering the implication of SFRP4 in early-onset colon cancer,particularly in the context of EMT,tumor metastasis,and invasion,and the effect of increased expression on colon cancer patient survival,SFRP4 might be a potential biomarker for early-onset colon cancer that could be targeted for diagnosis and/or disease therapy.

Model acupuncture point:Bone marrow-derived stromal stem cells are moved by a weak electromagnetic field

AIM To show the existence of a structural formative role of magnetic fields(MFs) with respect to biological objects by using our proposed model of an acupoint.METHODS We introduced a magnetised 10-100 μT metal rod(needle) into culture dishes with a negatively charged working surface and observed during 24 h how cells were arranged by MFs and by electrical fields(EFs) when attached. Rat and human bone marrow-derived stromal stem cells(r BMSCs and h BMSCs), human nonadherent mononuclear blood cells, NCTCs and A172 cells, and Escherichia coli(E. coli) were evaluated. The dish containing BMSCs was defined as the model of an acupoint. r BMSCs proliferative activity affected by the needle was investigated. For investigating electromagnetic field structures, we used the gas discharge visualisation(GDV) method.RESULTS During 24 h of incubation in 50-mm culture dishes, BMSCs or the nonadherent cells accumulated into a central heap in each dish. BMSCs formed a torus(central ring) with an inner diameter of approximately10 mm only upon the introduction of the needle in the centre of the dish. The cells did not show these effects in 35- or 90-mm culture dishes or hydrophobic dishes or rectangular cuvettes. NCTCs and A172 cells showed unstable the effects and only up to two weeks after thawing. Moreover, we observed that the appearance of these effects depended on the season. In winter, BMSCs showed no the effects. GDV experiments revealed that the resonant annular illumination gradually formed from 10 to 18-20 s in polar solutions with and without cell suspension of BMSCs, NCTCs and E. coli when using circular 50-mm dishes, stimulation at 115 V and switching of the electrode poles at 1 kH z. All these data demonstrate the resonant nature of the central ring. Significant influence of MFs on the rB MSC proliferation rate was not observed.CONCLUSION BMSCs can be moved by MFs when in the presence of a constant EF and MF, when the cells are in the responsive functional state, and when there is a resonant relationship between them.

Correlation of SFRP2 and TPX2 expression with cancer cell apoptosis and EMT process in cervical cancer

Objective: To study the correlation of SFRP2 and TPX2 expression with cancer cell apoptosis and EMT process in cervical cancer. Methods: Patients with cervical cancer who received surgical resection in No.174 Hospital of PLA between June 2015 and December 2016 were selected as the research subjects, moderate amount of cervical cancer lesion and adjacent lesion were collected after surgical resection to extract RNA, and then fluorescent quantitative PCR kit was used to determine the expression of SFRP2, TPX2, apoptosis genes and EMT genes in lesion tissue. Results: SFRP2, SARI, AIF, FHIT, NDRG4, MCPH1 and E-cadherin mRNA expression in cervical cancer lesion were greatly lower than those in adjacent lesions whereas TPX2, SP2, CyclinD1, Piwil2, HERC4, EFEMP1, EZH2, TGF-β1, N-cadherin and Vimentin1 mRNA expression were greatly higher than those in adjacent lesions;SFRP2 mRNA expression in cervical cancer lesion was positively correlated with SARI, AIF, FHIT, NDRG4, MCPH1 and E-cadherin mRNA expression, and negatively correlated with SP2, CyclinD1, Piwil2, HERC4, EFEMP1, EZH2, TGF-β1, N-cadherin and Vimentin1 mRNA expression whereas TPX2 mRNA expression was negatively correlated with SARI, AIF, FHIT, NDRG4, MCPH1 and E-cadherin mRNA expression, and positively correlated with SP2, CyclinD1, Piwil2, HERC4, EFEMP1, EZH2, TGF-β1, N-cadherin and Vimentin1 mRNA expression. Conclusion: The lowly expressed SFRP2 and highly expressed TPX2 in cervical cancer lesions can inhibit the apoptosis of cancer cells and promote the EMT process of cancer cells.

MiR-1180 from bone marrow-derived mesenchymal stem cells induces glycolysis and chemoresistance in ovarian cancer cells by upregulating the Wnt signaling pathway

Background:Bone marrow-derived mesenchymal stem cells(BM-MSCs)play an important role in cancer development and progression.However,the mechanism by which they enhance the chemoresistance of ovarian cancer is unknown.Methods:Conditioned media of BM-MSCs(BM-MSC-CM)were analyzed using a technique based on microRNA arrays.The most highly expressed microRNAs were selected for testing their effects on glycolysis and chemoresistance in SKOV3 and COC1 ovarian cancer cells.The targeted gene and related signaling pathway were investigated using in silico analysis and in vitro cancer cell models.Kaplan-Merier survival analysis was performed on a population of 59 patients enrolled to analyze the clinical significance of microRNA findings in the prognosis of ovarian cancer.Results:MiR-1180 was the most abundant microRNA detected in BM-MSC-CM,which simultaneously induces glycolysis and chemoresistance(against cisplatin)in ovarian cancer cells.The secreted frizzled-related protein 1(SFRP1)gene was identified as a major target of miR-1180.The overexpression of miR-1180 led to the activation of Wnt signaling and its downstream components,namely Wnt5 a,β-catenin,c-Myc,and CyclinD1,which are responsible for glycolysis-induced chemoresistance.The miR-1180 level was inversely correlated with SFRP1 mRNA expression in ovarian cancer tissue.The overexpressed mi R-1180 was associated with a poor prognosis for the long-term(96-month)survival of ovarian cancer patients.Conclusions:BM-MSCs enhance the chemoresistance of ovarian cancer by releasing miR-1180.The released miR-1180 activates the Wnt signaling pathway in cancer cells by targeting SFRP1.The enhanced Wnt signaling upregulates the glycolytic level(i.e.Warburg effect),which reinforces the chemoresistance property of ovarian cancer cells.