Advances in the Study of Chemokine-like Factor Superfamily Members in Tumors / 中国肺癌杂志

Chemokine-like factor-like MARVEL transmembrane domain containing member/chemokine-like factor superfamily member (CMTM/CKLFSF) including CKLF and CMTM1-CMTM8 are a new family of proteins linking chemokines and transmembrane superfamilies. CMTM not only have broad chemotactic activities, but also associate with hematopoietic system, immune system, and tumor development and metastasis closely. CMTM proteins are involved in key biological processes of cancer development, which include activation and recycling of growth factor receptors, cell proliferation and metastasis, and regulation of the tumor immune microenvironment. This is a new focus of research on the relationship between CMTM and tumors, because CMTM4/CMTM6 can be considered as a regulator for programmed cell death ligand 1 (PD-L1). This paper reviews the role of CMTM family members on cancer, especially in tumor growth, metastasis and immune escape, summarize the latest findings on the relationship between CMTM and non-small cell lung cancer, and explores the potential clinical value of CMTM as a novel drug target or biomarker.
.

Molecular Mechanism and Potential Clinical Value of CMTMFamily Members in Tumor Cells / 中国生物化学与分子生物学报

Nowadays, malignant tumor has become an important killer endangering human health, and the research on tumor has become a hot spot in the medical field. The CKLF-like MARVEL transmembrane domain containing family of genes (CMTM family), previously called chemokine-like factor super family (CKLFSF). CMTM family is a novel gene family firstly discovered by Chinese scholars and has a wide range of biological functions, which is closely related to human immune, hematopoietic and reproductive systems as well as tumors. CMTM family is dysregulated in many common tumors, can participate in a variety of signaling pathways closely related to the occurrence and development of cancer, which is closely related to the biological behavior changes of tumor cells. CMTM family members play an important role in the signaling molecular mechanisms that regulate the biological function of tumor cells and are able to regulate the signaling pathways of the tumor cell cycle and apoptosis. Including epithelial-mesenchymal transition process, epidermal growth factor receptor related signaling pathways, and closely related to the tumor immune microenvironment by regulating programmed death receptor-ligand 1. In this paper, we review the signaling molecular mechanisms of CMTM family in the biological processes of tumor cell proliferation, migration, invasion and apoptosis, as well as the potential clinical applications of CMTM family in ovarian cancer, breast cancer, liver cancer, gastric cancer, colorectal cancer, pancreatic cancer and other common malignant tumors. At present, the detailed molecular mechanism ofCMTM family related to tumor has not been fully clarified. Further study of the expression, biological role and regulatory mechanism of CMTM family in tumor will provide new ideas and targets for tumordiagnosis, treatment and prognosis prediction.

Association between CMTM5 gene and coronary artery disease and the relative mechanism / 北京大学学报(医学版)

OBJECTIVE@#To elucidate the correlation between CKLF-like MARVEL transmembrane domain containing member 5 (CMTM5) gene and the risk of coronary artery disease (CAD), and to detect the effects of CMTM5 gene expression changes on the ability of adhesion and migration of THP-1 cells.@*METHODS@#Using case-control method, a total of 700 hospitalized patients in Shijitan Hospital were enrolled in this study. CAD were diagnosed by coronary angiography, which was defined as at least one blood vessel diameter stenosis ≥50% according to the result of coronary angiography. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect CMTM5 gene expression; enzyme linked immunosorbent assay (ELISA) method to detect the plasma level of CMTM5; and Logistic regression to analyze CMTM5 genes and the risk of CAD. Human vascular endothelial cells (ECs) and THP-1 cells were cultivated, adhesion and Transwells experiments were used to evaluate the chemotactic capabi-lity of CMTM5 gene on THP-1 cells.@*RESULTS@#In this study, 350 CAD patients matched with 350 control patients were included. RT-PCR results revealed CMTM5 mRNA expression in CAD group was 3.45 times compared with control group, which was significantly higher than that in control group (P < 0.05). The levels of CMTM5 plasma protein in CAD group was (206.1±26.9) μg/L, which was significantly higher than that in control group (125.3±15.2) μg/L (P < 0.05). After adjusted for the risk factors of age, gender, BMI, smoking, hypertension, diabetes and hyperlipidemia, Logistic regression analysis results indicated that CMTM5 was the susceptibility factors of CAD, which still had significant correlation with CAD (P < 0.05). Adhesion and Transwells experiments results revealed that the numbers of adhesion and migration of THP-1 cells in CMTM5 overexpression ECs group (EO group) were significantly higher than that in lenti-mock infected ECs group (EO-MOCK group), non-infected ECs group (EN group), lenti-mock infected ECs group (ES-MOCK group), and CMTM5 suppression ECs group (ES group). On the contrary, the numbers of adhesion and migration of THP-1 cells in ES group were significantly lower than that in the other four groups (P < 0.01).@*CONCLUSION@#CMTM5 gene was closely related to the development of CAD. CMTM5 overexpression promoted the adhesion and migration of THP-1, which might play a part in the mechanisms of atherosclerosis and CAD.

Expression of CMTM6 in breast cancer tissues and its correlation with patients’ clinicopathological characteristics and prognosis / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the expression of chemokine-like factor-like MARVEL transmembrane domain-containing family member 6 (CMTM6) in breast cancer tissues and its correlation with clinicopathological features and prognosis of patients. Methods:Atotal of 136 breast cancer tissue chips (purchased from Superchip Company), including 42 pairs of matched cancer and paracancerous tissues, were used for this study. The expression level of CMTM6 in cancer and paracancerous tissues was detected by immunohistochemistry. The comparison of CMTM6 expression between breast cancer and paracancerous tissues was conducted by paired χ2 test. The relationship between CMTM6 expression in breast cancer tissues and the clinicopathological characteristics of patients was analyzed by χ2 test. Kaplan-Meier and Log rank test analyses were used to analyze the relationship between CMTM6 expression and the survival of patients, and Cox model was used to evaluate the effect of different indicators on the prognosis of patients. Results: The expression of CMTM6 in breast cancer tissues was significantly higher than that in paracancerous tissues (P<0.01). The expression of CMTM6 was correlated with pathological type of breast cancer and HER2 positivity (P<0.05). The survival time of patients in CMTM6 high expression group was significantly shorter than that of patients in CMTM6 low expression group (P<0.05). Pathological type (HR=10.374, 95%CI: 3.529-30.497, P<0.01), TNM stage (HR=4.599, 95%CI: 1.784-11.856, P<0.01), triple-negative breast cancer (HR=3.370, 95%CI: 1.055-10.761, P<0.05) and high expression of CMTM6 (HR=0.195, 95%CI: 0.073-0.518, P<0.01) were independent risk factors for prognosis of breast cancer patients. Conclusion: CMTM6 is highly expressed in breast cancer tissues, which can be used as a risk factor for prognosis evaluation of breast cancer patients.

Expression and clinical significance of CMTM6 in lung adenocarcinoma tissues / 中国肿瘤生物治疗杂志

@# Objective: To evaluate the expression of CKLF-like MARVEL transmembrane domain containing member 6 (CMTM6) in lung adenocarcinoma tissues, and to explore its correlation with the clinicopathologic features and prognosis of patients. Methods: Eighty-six pairs of cancer tissues and para-cancer tissues from patients that pathologically confirmed with lung adenocarcinoma were collected during September 2004 and June 2009 at the Third Affiliated Hospital of Soochow University. The expression levels of CMTM6 in above mentioned tissues were detected by immunohistochemistry. Serum of 52 patients with confirmed lung adenocarcinoma was collected before and after surgery, and serum of 32 healthy subjects was also collected. The levels of CMTM6 and PD-L1 in peripheral blood before and after surgery were measured and analyzed by ELISA. Chi-square test was used to analyze the relationship between CMTM6 expression and clinicopathological features; Kaplan-Meier method and Log-Rank test were used to analyze the survival data of patients. Results: CMTM6 was widely expressed in lung adenocarcinoma tissues; 30% of the tumor tissues showed an up-regulation as compared with para-cancer tissues, and 70% showed no difference. CMTM6 expression was associated with clinical stage and distant metastasis (all P<0.05), but not significantly associated with age, gender, tumor size, and T stage (P>0.05). Kaplan-Meier survival analysis showed the survival rate of patients with high CMTM6 expression was significantly lower than those with stable expression (P=0.014), and among patients at stage Ⅲ, the survival rate of patients with high CMTM6 expression was significantly lower than those with stable CMTM6 expression (P=0.001). Cox regression model analysis of multiple factors showed CMTM6 expression was an independent risk factor for the prognosis of patients with lung adenocarcinoma. CMTM6 expression in pre-surgery serum, post-surgery serum and healthy donors’serum showed statistically significant differences (P<0.05), which was significantly correlated with tumor size and age of the patients. Spearman correlation analysis showed a significant correlation between serum CMTM6 and PD-L1 expression level (r=0.623, P<0.01). Conclusion: CMTM6 is an independent risk factor for the prognosis of lung adenocarcinoma patients. It plays an important role in the occurrence and development of lung adenocarcinoma and it is a potential tumor suppressor gene.

Expressions of CMTM6 and PD-L1 protein in glioma tissues and their correlation with clinicopathological characteristics of patients / 中国肿瘤生物治疗杂志

@# Objective: : To investigate the expressions of chemokine-like factor superfamily 6 (CMTM6) and programmed cell death ligand 1 (PD-L1) in glioma tissues and their correlation with clinicopathological features of patients. Methods： ：From January 2012 to December 2015, 86 brain glioma tissues and 30 brain tissues (Control group) from patients operated with decompressive of craniotomy were collected from the FifthAffiliated Hospital of Zhengzhou University. The distribution and expressions of CMTM6 and PD-L1 protein in brain glioma tissues were detected by immunohistochemistry and WB methods. The differential expression of CMTM6 and PDL1 between glioma tissues and normal brain tissues was analyzed by t test of two independent samples. Single variant χ2 test was used to analyze the relationship between the expression of CMTM6, PD-L1 and the clinicopathological features of patients. Results: The expression of CMTM6 in glioma tissues was significantly higher than that in control tissues (P<0.01). The expression levels of CMTM6 and PD-L1 in high pathological grade (WHO III-IV) glioma tissues were significantly higher than those in low pathological grade (WHO I-II) glioma tissues (all P<0.01). The expression of CMTM6 was correlated with pathological grade, dizziness history, epilepsy seizure and PD-L1 expression (all P<0.05), while the expression of PD-L1 was correlated with pathological grade, epilepsy seizure and CMTM6 expression (all P<0.05). Conclusion: There is a correlation between the expression of CMTM6 and PD-L1 in glioma tissues, both of which are highly expressed and are expected to be used to study glioma signaling pathways.

Effect of microRNA-448 inhibitor on the proliferation and migration of human prostate cancer cells / 临床与实验病理学杂志

Purpose To observe the effect of specific miR-448 inhibitor on the proliferation and migration of prostate cancer cells and its molecular mechanism. Methods Real-time fluo-rescent quantitative polymerase chain reaction ( qRT-PCR) was used to detect the expression of miR-448 in normal prostate epi-thelial cells and cancer cells. The cells with the highest expres-sion of miR-448 were selected for follow-up experiment. The miR-448 inhibitor or miR-NC was transferred into prostate canc-er cells using liposome transfection reagent. The expression of miR-448 and CMTM3 mRNA were detected by qRT-PCR. The expression of related proteins were detected by Western blotting. MTT assay was used to detect the cell proliferation activity. Tr-answell assay was used to detect the cell migration ability. Re-sults The expression of miR-448 in normal prostate epithelial cells was significantly lower than that in cancer cells ( P ＜0. 01), and the expression of miR-448 was the highest in DU- 145 cells ( P ＜0. 01). The expression of miR-448 in DU-145 cells was down-regulated 48 h after transfection with miR-448 in-hibitor (P＜0. 01). The expression of CMTM3 mRNA was up-regulated (P＜0. 01). The expression of CMTM3, E-cadherin and β-catenin proteins were up-regulated. The expression of N-cadherin and Snail proteins were down-regulated. Cell prolifera-tion was decreased (P＜0. 05). Cell migration ability decreased (P ＜ 0. 01 ). Conclusion miR-448 is highly expressed in prostate cancer cells. miR-448 inhibitor can down-regulate the expression of miR-448 in DU-145 cells, up-regulate the expres-sion of CMTM3 protein, inhibit the proliferation and migration of prostate cancer cells, which showing a potential function in mo-lecular therapy of prostate cancer.

Expression and Significance of CMTM3 in Helicobacter pylori Infected Chronic Gastritis / 胃肠病学

Background: CKLF-like MARVEL transmembrane domain containing (CMTM) superfamily is involved in the occurrence and development of inflammation, cancer and a variety of diseases.Human CMTM3 has been proposed as a putative tumor suppressor gene.Aims: To investigate the expression of CMTM3 in Helicobacter pylori (Hp) infection-related chronic gastritis and its significance.Methods: Sixty cases of outpatients with chronic gastritis (30 Hp-positive and 30 Hp-negative) were enrolled for detection of CMTM3 and interleukin-6 (IL-6) expressions in gastric mucosa by immunohistochemistry.The correlation of expressions of CMTM3 and IL-6 was analyzed.Results: The positivity rates of CMTM3 and IL-6 in gastric mucosa were significantly higher in Hp-positive chronic gastritis than in Hp-negative ones (CMTM3: 63.3% vs.30.0%, P<0.05;IL-6: 73.3% vs.13.3%, P<0.01).In patients with Hp-positive chronic gastritis, CMTM3 and IL-6 were co-expressed in 53.3% (16/30) of the patients and localized in the same position.Expression of CMTM3 was positively correlated with IL-6 expression in Hp-positive chronic gastritis patients (r=0.58, P<0.05).Conclusions: CMTM3 is highly expressed in chronic gastritis patients with Hp infection.It may participate in the occurrence and development of Hp infection-related chronic gastritis with inflammatory cytokines such as IL-6.Hp infection might be one of the mechanisms involved in CMTM3 up-regulation.

Expression and localization of transmembrane protein CMTM2 in human testis and sperm / 北京大学学报(医学版)

Objective: To study the expression of transmembrane protein CMTM2 in the testis and sperm of adult males and to approach the potential function of the protein in the male reproductive system.Methods: The expression of CMTM2 in human testis and sperm was confirmed by Western blot.Immunohistochemical staining was used for detecting CMTM2 localization in the testis tissue, TRITC-CMTM2 and FITC-Hoechst double immunofluorescence staining was performed to examine the subcellular localization of CMTM2 in the human sperm before and after acrosome reaction, that is, immunofluorescent staining was used for detecting CMTM2 localization in both the testis and sperm before and after the acrosome reaction.Results: CMTM2 was presented in both human testis and sperm.In the testis, CMTM2 immunoreactive particles were observed mainly in the membrane of the different stages of spermatogenic cells.In the human sperm, its immunoreactivity was restrictively localized to the posterior head where sperm-egg fusion occurred, and the CMTM2 localization was not affected by sperm acrosome reaction.CMTM2 was widely expressed in seminiferous tubules of the human testis, mainly in the cell membranes of spermatogenic cells, which was consistent with the previous reports.The immunofluorescence performed on frozen human testis slides showed similar findings with immunohistochemistry, which gave weight to the localization of CMTM2 in the cell membranes of spermatogenic cells at different stages.TRITC-CMTM2 and FITC-Hoechst double immunofluorescence staining was performed to examine the subcellular localization of CMTM2 in the human sperm before and after acrosome reaction.CMTM2 was localized at the posterior head of sperm before and after acrosome reaction.The localization and expression of CMTM2 were not affected by sperm acrosome reaction.Conclusion: Expression of CMTM2 in the male reproductive system of the adult human exhibits cell-and region-specific patterns, which suggests that they may play an important role in spermatogenesis and sperm-egg fusion.The expression of CMTM2 in the male reproductive system of the adult human exhibits cell-and region-specific patterns, which suggests that they may play an important role in spermatogenesis and sperm-egg fusion.However, it still remains to be further elucidated about the definite role of CMTM2 in male reproductive system and the process of spermatogenesis.And in vitro fertilization experiments are needed to confirm the role of CMTM2 in fertilization in future.

Knockdown of CMTM3 promotes migration and invasion of PC3 cell in vitro / 北京大学学报(医学版)

Objective:To investigate the change of biological characteristics after stable knockdown of CKLF-like MARVEL transmembrane domain containing 3 (CMTM3)expression in PC3 by lentivirus shRNA and to reveal new therapeutic targets.Methods:The research includes two groups:sh393 is the experimental group in which CMTM3 is knocked down in PC3 cell line;shN is the control group in which CMTM3 is negatively knocked down.The expression of CMTM3 was detected by Western blot.The mi-gration ability of PC3 after stable knockdown was detected by Transwell and Wound healing assay.The invasion ability of PC3 was detected by Matrigel assay.Results were obtained from at least three indivi-dual experiments.Results:The expression of CMTM3 in sh393 group is significant lower than shN group (0.004 0 ±0.000 4 vs.0.490 0 ±0.055 7,P <0.001)detected by Western blot.It also had statistical significance in Matrigel assays (248.6 ±4.5 vs.113.0 ±3.3),Transwell (203.6 ±1.9 vs.103.0 ± 1.2)and Wound healing assays (95.0 ±2.9 vs.33.0 ±1.5)that knockdown of CMTM3 promoted mi-gration,and invasion of PC3 cells in vitro (P <0.001).Conclusion:Negative correlation exists between the stable knockdown of CMTM3 and change of biological characteristics in PC3 cells,and knocking down CMTM3 affects migration,and invasion ability in PC3 cells.

Effects of CMTM5 on EG-VEGF in prostate cancer cells / 中华内分泌外科杂志

Objective To investigate the effect of CKLF-like MARVEL transmembrane domain contai-ning member 5(CMTM 5)on EG-VEGF expression in prostate cancer cells , and to detect the potential mechanism of CMTM5 inhibiting prostate cancer .Methods The relative expression of CMTM 5 and EG-VEGF in prostate cells and prostate cancer cells was detected .Prostate cancer cells were given Plasmid transfection to overexpress CMTM5 and EG-VEGF expression was again detected .Then CMTM5 and EG-VEGF were compared between be-fore and after CMTM5 plasmid transfecting prostate cancer cells .Results Compared with the relative expresion of EG-VEGF and CMTM5 in prostate cells , prostate cancer cells showed high expression of EG-VEGF and low ex-pression of CMTM5, which were statistically significant , P<0.05.Compared with prostate cancer cells , the rela-tive expression of CMTM5 were obviously upregulated and EG-VEGF obviously decreased in prostate cancer cell after transfected by CMTM5 plasmid, which were statistically significant , P<0.05.Conclusions Prostate canc-er cells shows higher EG-VEGF expression and lower CMTM 5 campared with normal prostate cells .EG-VEGF is suppressed significantly when the prostate cancer cell is transfected by CMTM 5 plasmid and shows highlevel of CMTM5 expression , suggesting high expression of CMTM 5 may inhibit the development of prostate cancer by downregulating EG-VEGF expression .

Association between CMTM5 gene rs723840 single nucleotide polymorphism and high on asprin platelet reactivity / 北京大学学报(医学版)

Objective: To elucidate the correlation between the single nucleotide polymorphism of CKLF-like MARVEL transmembrane member 5 ( CMTM5 ) gene rs723840 and the occurrence of high on aspirin platelet reactivity ( HAPR) . Methods:The present study is a case-control study. A total of 210 hospitalized patients in Peking University First Hospital were enrolled. Aspirin response was assessed by 0. 5 g/L arachidonic acid (AA)-induced platelet aggregation ratio (PR), and ≥3/4 quartile of PR of the population was defined as HAPR. Accordingly all the enrolled 210 coronary artery diseases ( CAD) patients were divided into HAPR group and No-HAPR group. The genotypes were determined by poly-merase chain reaction ( PCR) and sequencing analysis for rs723840 of CMTM5 gene. Results:The geno-type frequencies in rs723840 C>T of CMTM5 gene conformed well to the Hardy-Weinberg equilibrium in both HAPR group and No-HAPR group. Between the two groups, the genotypes frequencies in HAPR and No-HAPR groups were 48 . 4%, 51 . 6%, 0 . 0% and 73 . 7%, 22 . 9%, 0 . 034%, respectively ( P=0. 004). The C, T allele frequencies were significantly different in the two groups (P =0. 031,OR =0 . 501 , 95%CI:0 . 264-0 . 947 ) . Conclusion:Our study finds a significant correlation between CMTM5 gene rs723840 polymorphism and high on aspirin platelet reactivity.

The expression and role of CMTM family in tumor / 国际肿瘤学杂志

CMTM family is silenced and down-regulated in several kinds of tumors.Its aberrant expression has a strong association with the development,progression and metastasis of tumors.Thus,CMTM family is potential tumor suppressor genes.Epigenetics mechanism is the essential mechanism of the aberrant expression in this gene family.The discovery of this research gives a new direction to the clinical treatment of tumor.

CMTM1-v17, a new potential corepressor of androgen receptor / 北京大学学报(医学版)

Objective: To analyze the expression of CMTM1-v17 in normal prostate tissue and prostate carcinoma originated cell lines, and study its impact on the transactivation of androgen receptor and the possible mechanism. Methods: The expression of CMTM1-v17 in normal prostate tissue was analyzed with immunohistochemistry method. In immounocytochemistry was used to analyze the expression of CMTM1-v17 in prostate carcinoma originated cell lines. Luciferase assay was used to study the impact of CMTM1-v17 on the transactivation of AR and its mechanism. Results: The results of immunohistochemistry showed that CMTM1-v17 was highly expressed in prostate. In prostate cancer originated cell lines, CMTM1-v17 could also be detected in prostate cancer originated cell lines PC3, Du145 and LNCaP. And the results of luciferase implied that the relative luciferase activity of the PC3 cells transfected with 1 ?g and 2 ?g pCDI-CMTM1-v17 plasmids separately were 70.8 and 34.7, compared with the control set as 100. When trichostatin A, the inhibitor for histone deacetylase, was used, the repression of androgen receptor could be recovered with trichostatin A treatment,for the relative luciferase activity of the PC3 cells transfected with 1 ?g and 2 ?g pCDI-CMTM1-v17 plasmids and treated with 100 nmol/L trichostatin A rebound to 90.9 and 86.4. Conclusion: CMTM1-v17 is highly expressed in both normal prostate and prostate carcinoma originated cell lines. It may recruit histone deacetylas to inhibit the function of androgen receptor.

CMTM5 inhibits the tumor cell behavior of prostate cancer by downregulation of HER2 / 北京大学学报(医学版)

Objective:To discover whether there is any other pathway than EGFR which has interaction with HER2 that makes a different down stream on cancer manners and whether CMTM5 plays a role in HER2 related tumor performance.Methods:HER2 and CMTM5 were detected on prostate cancer tissue chip using IHC.The protein levels of HER2、Cyclin D1 and CMTM5 were compared had with or without CMTM5 plasmid transfection on PC3 cell line to insure their relationship.Results:The author demonstrated that the HER2 had been up regulated in prostate cancer epithelium while CMTM5 down regulated.Overexpression of CMTM5 in PC3 could lower the HER2 and Cyclin D1 protein level.Conclusion:The results suggest that in prostate cancer HER2 has a different partner in the signaling transduction other than EGFR as in a traditional pathway.CMTM5 may have a chance to be chosen as the next potential treatment bio-target of prostate cancer.

Preparation and identification of polyclonal antibody against CMTM1-v17/CKLFSF-v17 / 中国免疫学杂志

Objective:To analyze a novel antigenic determinant of Homo sapiens CMTM1-v17 protein and prepare polyclonal antibody.Methods:By means of DNA recombination method, the C-terminal(The coding sequences of 118-149 residues) of CMTM1-v17 gene was inserted into pGEX-4T-2 vector. The recombinant plasmid was transformed into E.coli XL-1 Blue to express the fusion protein of GST/CMTM1-v17. The GST tag of the purified fusion protein was removed by enzymatic cleavage with thrombin. The polyclonal anti-CMTM1-v17 antibody were prepared by immunizing rabbits with the purified protein. Highly-specific, polyclonal anti-CMTM1-v17 antibodies was purified by affinity chromatographic column matrix coupled with antigen. The specificity of the anti-CMTM1-v17 antibody was identified by Western blot and immunohistochemistry.Results:The polyclonal antibody showed specific response to CMTM1-v17 by Western blot analysis and immunohistochemistry. These results suggested that the polyclonal antibody can be used for further studies of CMTM1-v17 function.Conclusion:A polyclonal anti-CMTM1-v17 antibody has been synthesized successfully.