Role of Autophagy-apoptosis Balance in Colorectal Cancer and Traditional Chinese Medicine Intervention： A Review / 中国实验方剂学杂志

Colorectal cancer （CRC）， a malignant tumor of the digestive system， originates from the colorectal mucosa epithelium and is usually asymptomatic until it progresses to an advanced stage. With high incidence around the globe and the increasingly younger patients， this disease poses a serious threat to the health and lives of the patients. Although the pathogenesis of this disease is not fully understood， it is generally believed that it is associated with autophagy， apoptosis， and inflammation. Autophagy and apoptosis as two types of programmed cell death are subject to complex interactive regulation， and the imbalance between them is closely related to the occurrence， development， and prognosis of a variety of diseases. Studies have shown that autophagy-apoptosis balance plays a key role in CRC. On the one hand， autophagy and apoptosis coordinate with each other to inhibit CRC cell growth. On the other hand， autophagy can antagonize apoptosis to promote CRC cell growth. In clinical practice， surgery is often combined with radiotherapy and chemotherapy to treat CRC， which can control the progression of CRC to a certain extent but has serious adverse effects and poor long-term results. In recent years， traditional Chinese medicine （TCM） has been proved to be effective in the treatment of CRC. Studies have shown that numerous herbal active components can promote CRC cell death by regulating the autophagy-apoptosis balance， thereby blocking the progression of this disease. The process of autophagy-apoptosis balance in regulating cell activities has similar theoretical connotations with the Yin and Yang theory of TCM. Applying TCM in regulating autophagy-apoptosis balance at various stages of CRC has become a frontier， while the comprehensive elaboration remains to be conducted. By reviewing the relevant studies in recent years， this paper introduces the correlation between the Yin and Yang theory and the autophagy-apoptosis balance， the role of autophagy-apoptosis balance in CRC， and the research progress in the application of 27 Chinese herbal active components such as flavonoids， terpenoids， glycosides， and phenols capable of regulating autophagy-apoptosis balance in the treatment of CRC. The active components in Chinese medicines can recover the autophagy-apoptosis balance in CRC by acting on microtuble-associated protein 1 light chain 3（LC3）， Beclin-1， and B-cell lymphoma-2（Bcl-2）to regulate multiple signaling pathways such as protein kinase B（Akt）/mammalian target of rapamycin（mTOR）and reavtive oxygen species（ROS）/ c-Jun N-terminal kinase（JNK）， thus balancing Yin and Yang. This review aims to provide a reference for the treatment of CRC and the development of new drugs.

CXCL8 Promotes M2 Macrophage Chemotaxis and Infiltration in Colorectal Cancer Microenvironment / 中国生物化学与分子生物学报

CXC chemokine ligand 8 (CXCL8) is highly expressed in many human tumors including colorectal cancer, and it can promote the malignant progression of tumors. It was reported that M2 macrophages were abundant in colorectal cancer microenvironment, but whether CXCL8 affects the infiltration of M2 macrophages and its potential mechanism are not yet clear. The study aimed to investigate the effect of CXCL8 on M2 macrophage infiltration and chemotaxis in the colorectal cancer. Firstly, we analyzed the CXCL8 expression and immune cell infiltration in human colorectal cancer tissues from TCGA RNA-seq data. The expression of CXCL8 was verified by immunohistochemistry in tissues obtained from Shanxi Provincial Cancer Hospital. Then, Western blot and qRT-PCR were employed to detect CXCL8 expression in five colorectal cancer cell lines. THP-1 cells were allowed to differentiate into M2 macrophages via the phorbol myristate acetate (PMA) and IL-4 treatment, followed by detection of the chemotaxis of M2 macrophages towards HCT116, SW480 and CXCL8-HCT116, CXCL8-SW480 cell lines. HCT116 and SW480 cells were treated with interleukin 1β (IL-1β) to detect the expression of CXCL8, and co-cultured with M2 macrophages to analyze the chemotactic activity. The results revealed that the expression of CXCL8 was increased in pairs of CRC tissues versus normal adjacent tissues, and there were more M2 macrophage infiltration in cancer tissues with high expression of CXCL8. The mRNA and protein expression of CXCL8 in HCT116 and SW480 were increased after the IL-1β treatment (P < 0. 05). We confirmed that CXCL8 is a chemotactic factor for M2 macrophages by transwell assays (P<0. 05). In conclusion, CXCL8 in colorectal cancer cells can be induced by IL-1β in colorectal cancer cells and the upregulation of CXCL8 can promote the chemotaxis of M2 macrophages. The massive infiltration of M2 macrophages in colorectal cancer microenvironment may be related with the increased expression of CXCL8.

Siglec-15 Promotes the Proliferation and Migration of Colorectal Cancer Cells and Inhibits the Infiltration of CD4 / 中国生物化学与分子生物学报

Sialic acid binding immunoglobulin type lectin-15 (Siglec-15), one of the Siglec gene family members, is a new immunosuppressive molecule. Siglec-15 is highly expressed in a variety of human tumor cells and tumor-associated macrophages, but the biological function of Siglec-15 in colorectal cancer (CRC) and its effect on the tumor immune microenvironment remains poorly understood. This study aimed to investigate the effects of aberrant expression of Siglec-15 on the biological behaviors of CRC cells and the infiltration of CD4

Efficacy and Mechanism of Shenbai Jiedu Prescription Against Proliferation of HCT116 Cells / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism by which Shenbai Jiedu prescription （SBJDF） inhibits the proliferation of colorectal cancer （CRC） HCT116 cells. MethodAfter 48 h treatment of HCT116 cells with SBJDF （0， 0.25， 0.5， 1， 2， 4 g·L-1）， the viability of HCT116 cells were determined by methyl thiazolyl tetrazolium （MTT） colorimetry. Following the classification of cells into blank control group and SBJDF （1， 2， 4 g·L-1） groups， the effect of SBJDF on HCT116 cell morphology was observed under an inverted microscope. The effects of SBJDF on the proliferation of HCT116 cells and mitochondrial membrane potential （Δψm） were detected by colony formation assay and JC-1 probe， respectively. The flow cytometry was then performed for determining cell cycle distribution and apoptosis. The effects of SBJDF on cell cycle-， apoptosis-， and nuclear factor kappa-B （NF-κB） signaling pathway-related proteins were determined by Western blot. ResultSBJDF effectively inhibited the vitality of HCT116 cells and changed their morphology in a concentration-dependent manner. Compared with the blank control group， SBJDF at 1， 2， 4 g·L-1 significantly reduced cell colony formation （P<0.05， P<0.01），and SBJDF at 2 and 4 g·L-1 arrested the HCT116 cell cycle at G0/G1 phase （P<0.05， P<0.01）. Compared with the blank control group， SBJDF at 1， 2， 4 g·L-1 remarkably down-regulated the protein expression of CyclinD1 （P<0.05， P<0.01）. SBJDF at 2 and 4 g·L-1 lowered the CyclinA2 and cyclin-dependent kinase 4 （CDK4） （P<0.05， P<0.01）. SBJDF at 4 g·L-1 reduced the cyclin-dependent kinase 1 （CDK1） （P<0.01）. Compared with the blank control group， SBJDF at 2 and 4 g·L-1 induced HCT116 cell apoptosis， down-regulated the protein expression of anti-apoptosis-related proteins Bcl-2 and Bcl-xl as well as the NF-κB signaling pathway-related proteins IκB kinase α （IKKα），inhibitor α of NF-κB （IκBα），and phospho-NF-κB p65 （p-p65） （P<0.05， P<0.01）， and diminished the mitochondrial membrane potential of HCT116 cells. ConclusionSBJDF inhibits the proliferation of HCT116 cells， which may be related to its inhibition of the activation of NF-κB signaling pathway and the induction of cell cycle arrest and apoptosis.

Detección de mutaciones del Gen KRAS en pacientes paraguayos con cáncer colorrectal / Detection of KRAS Gene mutations in paraguayan patients with colorectal cancer

El estado mutacional del KRAS ha sido considerado como biomarcador para tratamientos biológicos tras varios ensayos clínicos realizados en pacientes con cáncer colorrectal metastásico. Reportes recientes indican que las frecuencias de mutación del gen KRAS en pacientes con CCR de Asia, Europa y Latinoamérica están entre el 24%, 36% y 40%, respectivamente. Paraguay no cuenta con este tipo de informes, a pesar de registrar anualmente en promedio 75 nuevos casos de pacientes diagnosticados con CCR sólo en el Servicio de Cirugía General del Hospital Central del Instituto de Previsión Social (IPS). El presente trabajo ha implementado este análisis de rutina, prerrequisito obligatorio para la administración de fármacos basados en anticuerpos terapéuticos, y revelado una frecuencia de mutación del gen KRAS del 34% en pacientes paraguayos con CCR que acuden a los Servicios del Hospital Central del IPS

The mutational status of the KRAS has been consider as a biomarker for biological treatments after several clinical trials carried out in patient with metastatic colorectal cancer. Recent reports indicate that the KRAS gene mutation frequencies in CRC patients from Asia, Europe, and Latin America are between 24%, 36%, and 40%, respectively. Paraguay does not have this kind of reports, despite registering an average of 75 new cases of patients diagnosed with CRC per year only in the General Surgery Service of the "Central Hospital - Instituto de Prevision Social (IPS)". The present work has implemented this routine analysis, a mandatory prerequisite for the administration of drugs based on therapeutic antibodies and revealed a KRAS gene mutation frequency of 34% in Paraguayan patients with CRC who attend the IPS Central Hospital Services

High expression of FABP4 in colorectal cancer and its clinical significance / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

OBJECTIVES@#To investigate the relationship between the fatty acid-binding protein 4 (FABP4) and colorectal cancer (CRC).@*METHODS@#Using an enzyme-linked immunosorbent assay (ELISA), we measured the expression of FABP4 in plasma of 50 patients who underwent surgery for CRC from October 2017 to May 2018 and 50 healthy controls. The content of the visceral fat area (VFA) as seen with abdominal computed tomography (CT) scanning was measured by ImageJ software. The expression levels of FABP4, E-cadherin, and Snail proteins in CRC and adjacent tissues were determined by immunohistochemistry.@*RESULTS@#The mean concentration of plasma FABP4 of CRC patients was higher than that of the control group (22.46 vs. 9.82 ng/mL; @*CONCLUSIONS@#High LPA and VFA were risk factors for increased plasma FABP4 in CRC patients. FABP4 protein was highly expressed in CRC tissues and associated with TNM stage, differentiation, and lymph node metastasis of CRC. The level of FABP4 in CRC tissue was correlated with E-cadherin and Snail expression, suggesting that FABP4 may promote CRC progression related to epithelial-mesenchymal transition (EMT).

Circular RNA PVT1 Promotes Proliferation and Represses Apoptosis of Colorectal Cancer Cells / 中国生物化学与分子生物学报

Circular RNAs (circRNAs) are a novel class of long-chain non-coding RNAs. Preceding evidence has showed that circRNAs participate in the development and progression of various tumors. In the present study, we investigated the expression of circRNAs in 5 paired colorectal cancer (CRC) tissues and adjacent normal tissues with circRNA high-throughput sequencing. Totally 477 differentially expressed circRNAs were identified between CRC tissues and non-cancerous matched tissues, which included 252 significantly overexpressed circRNAs and 225 downregulated circRNAs. CircRNA plasmacytoma variant translocation 1 (circPVT1), the most up-regulated expression circRNA, was further confirmed by qRT-PCR in 150 colorectal cancer tissues and matched normal mucosae. Our data revealed that circPVT1 showed a significant up-regulation trend in CRC tissues compared with matched normal mucosae. Similarly, compared with normal colorectal mucosa cells, the expression of circPVT1 in colorectal cancer cell lines was significantly up-regulated (P<0. 05). Functionally, silence with siRNA or overexpression of circPVT1 in colorectal cancer cells was applied to determine the biological functions of circPVT1, including cell proliferation, apoptosis, and cell cycle, etc. The results show that circPVT1 expression significantly attenuated apoptosis, induced replication and promoted proliferation of colorectal cancer cells in vitro. In summary, our findings indicate that circPVT1 plays an oncogenic role in CRC and might be a potential novel target for CRC therapy.

Expression of TAGLN in colorectal cancer tissues and its effect on malignant biological behaviors of SW480 cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the expression of transgelin (TAGLN) in colorectal cancer (CRC) tissues and its effect on the proliferation, migration and invasion of CRC SW480 cells. Methods: Surgically resected CRC tissues and corresponding para-cancerous tissues of 97 CRC patients from May 2015 to August 2016 in the Affiliated Tumor Hospital of Zhengzhou University were collected; In addition, CRC cell lines SW620, SW480, HCT116 and normal colorectal mucosal cell line FHC were also collected for this study. Immunohistochemical staining was used to detect the expression of TAGLN in CRC tissues, and the correlation between TAGLN and patients’ clinicopathological features was analyzed. Quantitative Real-time quantitative polymerase chain reaction (qPCR) and Western blotting (WB) were used to detect the mRNA and protein expressions of TAGLN in CRC cell lines. si-TAGLN and si-Ctrl were respectively transfected into SW480 cells by liposome transfection method. The effects of silencing TAGLN on the proliferation, migration and invasion of SW480 cells were detected by CCK-8, Wound-healing assay and Transwell assay, respectively; and the expression of EMT-related proteins E-cadherin, N-cadherin and vimentin were detected by WB. Results: The positive expression rate of TAGLN in CRC tissues was significantly higher than that in para-cancerous tissues (P<0.01), and TAGLN expression was correlated with TNM stage, degree of tumor differentiation and lymph node metastasis in CRC patients (P<0.05 or P<0.01). The mRNA and protein expression levels of TAGLN in SW480 cells were significantly higher than those in FHC cells (all P<0.01). After TAGLN silence, the proliferation, invasion and migration ability of SW480 cells were significantly reduced (all P<0.01), the expression level of E-cadherin in SW480 cells was increased, while the expression levels of N-cadherin and vimentin were decreased (all P<0.01). Conclusion: TAGLN is highly expressed in CRC tissues and cells. Silencing TAGLN can inhibit the proliferation, invasion and migration of CRC cells, suggesting that TAGLN plays an important role in the occurrence and development of CRC.

Expression of lnc-MTBP-5 in colorectal cancer and its effect on cell invasion / 上海交通大学学报（医学版）

Objective: To analyze the expression of lnc-MTBP-5 in colorectal cancer (CRC), and explore the effect of lnc-MTBP-5 on the invasion of CRC cells and its potential mechanism. Methods: Bioinformatics data from PRJNA218851 and PRJNA376161 data sets were extracted from the Sequence ReadArchive (SRA) database to screen CRC metastasis associated lncRNAs. The Cancer Genome Atlas (TCGA) was used to analyze the expression of lnc-MTBP-5 in CRC tissues and normal tissues, its relationship with the prognosis of patients, and its correlation with metastasis related factors. Quantitative real-time PCR (qPCR) was used to detect the expression of lnc-MTBP-5 in normal intestinal epithelial cells and CRC cells, as well as 53 CRC tissues and para-cancer mucosa. After lnc-MTBP-5 was down-regulated in CRC cells, CCK-8 assay, clone formation and Transwell assay were performed to observe the effect of lnc-MTBP-5 on the proliferation and invasion ability of CRC cells. Results: Lnc-MTBP-5 was associated with CRC metastasis. The expression of lnc-MTBP-5 was significantly increased in 5 CRC cell lines and CRC tissues. Compared with patients with low expression of lnc-MTBP-5, CRC patients with high expression of lnc-MTBP-5 were younger, had higher American Joint Committee on cancer (AJCC) staging, and were prone to metastasis. Lnc-MTBP-5 was positively correlated with CRC metastasis associated in colon cancer 1 (MACC1), mesenchymal to epithelial transition factor (MET) and cadherin-associated protein. After lnc-MTBP-5 was down-regulated, the invasion ability of CRC cells decreased. Conclusion: Lnc-MTBP-5 is up-regulated in CRC cell lines and CRC tissues, and it is negatively correlated with the prognosis of patients. Lnc-MTBP-5 can promote the invasion ability of CRC cells, which may be related to MACC1-HGF (hepatocyte growth factor)/MET pathway.

Effect of DUOX2 on sensitivity of colorectal cancer cells to 5-fluorouracil / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of double oxidase 2 (DUOX2) on the sensitivity of colorectal cancer (CRC) cells to 5-fluorouracil (5-FU). Methods: CRC cell lines DLD-1, SW480, HCT116, SW620 and normal intestinal epithelial cell line NCM460 were selected, and the expression of DUOX2 in these cell lines were detected by qPCR. DUOX2 expression in HT-29 and HCT116 cells was stably knocked down by lentivirus infection technique. The knockdown efficiency was detected by qPCR and WB. Cells in sh-Control and sh-DUOX2 groups were treated with 5-FU at different concentrations (0, 5, 10, 20, 40, 80, 120 μg/ml). The effects of 5-FU on cell proliferation, apoptosis and cell cycle were detected by CCK-8 method and flow cytometry. HT29 cell transplanted xenograft model in nude mice was constructed to observe the effect of DUOX2 gene on the treatment efficacy of 5-FU. Results: the expression level of DUOX2 mRNA in CRC cells was significantly higher than that in NCM460 cells (P<0.05 or P<0.01). Compared with sh-Control group, the mRNA and protein expressions of DUOX2 in sh-DUOX2 group were significantly decreased (all P<0.01); the sensitivity of cells to 5-FU was enhanced, the apoptosis rate and the ratio of cells at G0/G1 phase were significantly increased (all P<0.01), and the ratio of cells at G2 and S phase was significantly decreased (all P<0.01). There was no significant difference in tumor volume and mass between sh-Control group and sh-DUOX2 group without 5-FU treatment (all P>0.05), but the volume and mass of transplanted tumor in sh-DUOX2+5-FU group after 5-FU treatment was significantly lower than that in sh-Control+5-FU group (all P<0.01). Conclusion: The sensitivity of CRC cells to 5-FU can be significantly enhanced by knocking down DUOX2 gene.

Advances in treatment of peritoneal metastasis from colorectal cancer / 中国肿瘤临床

Peritoneum is the third common metastatic site of colorectal cancer (CRC) following liver and lung. CRC peritoneal metastasis (PM) has been reckoned as an advanced disease with dismal prognosis. With the development of modern chemotherapeutic modalities, the prognosis of patients with metastatic CRC has been dramatically improved, yet patients with CRC PM achieved few survival benefits. It is the emergence and combination of cytoreductive surgery and hyperthermic intraperitoneal chemotherapy that prolong the survival of PM cases. What's more, novel treatments, pressurized intraperitoneal aerosol chemotherapy, and intraperitoneal MOC31PE immunotoxin treatment, for instance, have been under investigation and preliminary results are promising. We aim to comprehensively review the literature focusing on the clinical treatment of PM.

Analysis of differentially expressed genes and signaling pathways in colorectal cancer with liver metastasis / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore the key genes and molecular mechanisms of liver metastasis in colorectal cancer (CRC), and to provide potential targets and biomarkers for the treatment of CRC with liver metastasis. Methods: Based on the bioinformatics method, the gene data sets of CRC liver metastasis were downloaded from the GEO database to screen the differentially expressed genes (DEGs); the GO and KEGG enrichment analyses of DEGs were performed by using DAVID online tool, and the protein-protein interaction (PPI) network was constructed to screen out the key genes, and subsequently the prognosis was analyzed. Results: A total of 321 DEGs were selected from 183 CRC specimens and 39 liver metastasis specimens, including 153 up-regulated genes and 168 downregulated genes. The results of enrichment analysis of GO and KEGG showed that the functions of DEGs were mainly related to protein activation cascade, inflammatory response, extracellular matrix, platelet degranulation, complement and coagulation cascade reaction etc. 8 key CRC genes (ALB, APOB, FGA, F2, APOA1, SERPINC1, FGG and AHSG) were screened by PPI network. Survival analysis showed that patients with high expressions of SERPINC1 and FGG had poor prognosis(all P<0.05). Conclusion: The biological functions and signaling pathways of DEGs are related to the occurrence and development of liver metastasis. The 8 key genes may be the potential therapeutic targets of CRC liver metastasis, and SERPINC1 and FGG may be new prognostic markers.

Treatment efficacy and safety of apatinib monotherapy in patients with failed standard regimen for advanced colorectal cancer / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the efficacy and safety of apatinib monotherapy in the treatment for patients with advanced colorectal cancer (CRC) who failed standard regimen. Methods: The required sample size in this prospective study was calculated with the PASS 15 software. A total of 52 patients with advanced colorectal cancer who failed standard regimen from July 2017 to August 2018 were included in this study. The patients were given apatinib monotherapy with an initial dosage of 750 mg or 500 mg. The objective remission rate (ORR) and disease control rate (DCR) were evaluated; the patients were followed up and progression-free survival (PFS) and overall survival (OS) were evaluated, and adverse events during treatment were recorded. The primary endpoint of this study was PFS, and secondary endpoints were ORR, DCR, OS and safety. Result: Of the 52 patients included, 45 patients, all of whom were late stage CRC patients with at least two systematic chemotherapeutic treatments, were available for efficacy evaluation. Treatment efficacy evaluation showed complete response of 0 case, partial response of 5 cases, stable disease of 30 cases and progression disease of 10 cases; the ORR was 11.11%, and the DCR was 77.78%. The prognosis data indicated that the median PFS of the 45 CRC patients was 3.95 months (95% CI=3.16-4.74), and the median OS was 10.3 months (95% CI=5.70-14.90). In terms of adverse events evaluation, the adverse reactions with grade 3 or above were hand-foot syndrome (6 cases, 13.33%), hypertension (5 cases, 11.11%), proteinuria (3 cases, 6.67%), diarrhea (3 cases, 6.67%), fatigue (2 cases, 4.44%) and bleeding (1 case, 2.22%). Conclusion: Apatinib monotherapy for patients with advanced colorectal cancer, who failed the standard regimens, has potential clinical benefits, and the overall toxicity profile is manageable.

EffectofGSTP1geneticvariationontherecurrenceriskandprognosisofcolorectal cancerpatients received postoperative adjuvant chemotherapy / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the influence of glutathione S-transferase P-1 (GSTP1) genetic variation on the recurrence risk and prognosis of colorectal cancer (CRC) patients received postoperative adjuvant chemotherapy. Methods: The clinical data of 195 CRC patients, who received postoperative adjuvant chemotherapy in the Department of Gastroenterology of the FirstAffiliated Hospital of Zhengzhou University from January 2010 to December 2018, were collected for this study. 5-fluorouracil (5-FU) based adjuvant chemotherapy was given after surgical resection. The recurrence status of the patients was assessed during hospitalization period, and the long-term survival data of patients were obtained by telephone follow-up after finishing the scheduled adjuvant chemotherapy. GSTP1 genotyping was performed with the DNA extracted from peripheral blood specimens, and its correlation with patients’clinical characteristics wasanalyzed.Additionally, RNAwasextractedfrom peripheral blood mononuclear cell (PBMC) specimens of some CRC patients that prior to chemotherapy for GSTP1 mRNA expression analysis. The univariate analysis of genotypes and prognosis was carried out by Kaplan-Meier survival analysis, and adjusted by multivariate Cox regression model. Results: The median disease-free survival (DFS) of the 195 CRC patients was 4.8 years, and the median overall survival (OS) was 6.2 years. Polymorphism analysis indicated that the I105VlocusofGSTP-1coding region was correlated with prognosis. The prevalence of I105V in the study population: AA genotype of 135 cases (69.23%), AG genotype of 56 cases (28.72%) and GG genotype of 4 cases (2.05%), the minor allele frequency of I105V was 0.16. The genotype distribution was in accordance with Hardy-Weinberg equilibrium (P>0.05). The analysis of recurrence risk and prognosis found that the median DFS of patients with AA genotype and AG/GG genotype was 5.7 and 3.9 years respectively (P<0.01), while the median OS of two groups of patients was 7.0 and 4.5 years respectively (P<0.01). The multivariate Cox regression results indicated that AG/GG genotype was an independent factor for OS (OR=1.54, P<0.05). The mRNA expression of GSTP1 in PBMC of the patients with AG/GG genotypes were significantly higher than those patients with AA genotype (P<0.01). Conclusion: GSTP1 I105V genetic variation influences the recurrence risk and prognosis of CRC patients received postoperative adjuvant chemotherapy possibly via mediating the mRNAexpression of GSTP1.

Association between genetic variation of kinase insert domain receptor and prognosis in colorectal cancer patients received 5-FU based adjuvant chemotherapy / 中国肿瘤生物治疗杂志

@# Objective: To investigate the association between genetic variation of kinase insert domain receptor (KDR) and the prognosis in colorectal cancer (CRC) patients received 5-FU based adjuvant chemotherapy. Methods: The clinical data of 176 CRC patients, who underwent surgical treatment at the Department of Anus and Intestine Surgery, People’s Hospital of Zhengzhou during January 2012 and December 2017, were retrospectively analyzed, and 93 cases of tumor tissues were collected for this study. The genotype of KDR polymorphism locus was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). qPCR was used to detect the expression of KDR mRNAin colorectal cancer tissues. The correlation between the polymorphism genotypes and other variables was analyzed by logistic regression model. The expression of different genotypes of KDR was analyzed by nonparametric test. The relationship between KDR genotype and prognosis of patients was analyzed by Kaplan-Meier survival analysis, and the other variables were adjusted by Cox risk scale model. Results: Of the polymorphisms analyzed, only rs2071559 was of clinical significance. The distribution frequency of KDR rs2071559 in 176 CRC patients was as follows: TT genotype in 95 cases (53.98%), TC genotype in 70 cases (39.77%) and CC genotype in 11 cases (6.25%); the minor allele frequency was 0.26; and the distribution of three genotypes was in accordance with Hardy-Weinberg's Equilibrium (P=0.690). The median disease free survival (mDFS) of patients carrying C allele and wild type TT genotype was 4.4 and 3.2 years, respectively (P<0.05); The median overall survival (mOS) of patients with TC/CC genotype and TT genotype was 5.2 and 4.0 years, respectively (P<0.05). After COX model modification, the effect of TC/CC genotype on mOS was still statistically significant (OR=0.55, P<0.05). The mRNA expression of KDR in cancer tissues of the patients with TC/CC genotypes were significantly lower than those of the wild type TT genotype (P<0.01). Conclusion: The polymorphism of KDR rs2071559 is associated with clinical outcomes in patients with colorectal cancer. KDR rs2071559 may affect the prognosis of colorectal cancer patients by affecting the mRNAexpression of KDR.

Altered intestinal microbiota associated with colorectal cancer / 医学前沿

The gut microbiota plays an important role in the development and progression of colorectal cancer (CRC). To learn more about the dysbiosis of carcinogenesis, we assessed alterations in gut microbiota in patients with CRC. A total of 23 subjects were enrolled in this study: 9 had CRC (CRC group) and 14 had normal colons (normal group). The microbiome of the mucosal-luminal interface of each subject was sampled and analyzed using 16S rRNA gene amplicon sequencing. We also used Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) to predict microbial functional profiles. The microbial composition of the mucosal lumen differed between the groups, and the presence of specific bacteria may serve as a potential biomarker for colorectal carcinogenesis. We identified a significant reduction in Eubacterium, which is a butyrate-producing genera of bacteria, and a significant increase in Devosia in the gut microbiota of CRC patients. Different levels of gut microflora in healthy and CRC samples were identified. The observed abundance of bacterial species belonging to Eubacterium and Devosia may serve as a promising biomarker for the early detection of CRC.

IL-18 over-expression inhibits proliferation of human colorectal cancer HCT-116 cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects of interleukin-18 over-expression on the in vitro and in vivo proliferation of human colorectal cancer (CRC) HCT-116 cells. Methods: A recombinant lentivirus vector containing human IL-18 gene fragment was constructed. Then theCRC HCT-116 cell line stably expressing human IL-18 (HCT-116/IL-18) was obtained by recombinant lentivirus transfection. In vitro proliferation of HCT-116/IL-18 cells and wild-type HCT-116 cells was determined by CCK-8 method. The expressions of IL-18, Cyclin D1, proliferating cell nuclear antigen (PCNA) and DNA damage repair enzyme (PARP) were detected by Western blotting. HCT-116 and HCT-116/IL-18 cells were inoculated into left and right axillas of Balb/c nude mice, respectively. Then the tumorigenicity and the growth of transplanted tumor were observed. The expressions of IL-18 and PCNAin xenograft tissues were detected by immunohistochemistry analysis. Results: IL-18 gene over-expression in HCT-116 cells could delay the proliferation of HCT-116 cells (P<0.05 or P<0.01). PARP expression was increased significantly and PCNA, Cyclin D1 expression were decreased in HCT-116/ IL-18 cells as compared to that of HCT-116 cells (P<0.01).The tumorigenicity of HCT-116/IL-18 cell was significantly decreased in nude mice with a tumor-formation rate of 43%; Compared with control group, HCT-116/IL-18cell line had a longer tumorigensis time, slower growth and smaller tumor volume; moreover, PCNAprotein expression was down-regulated in HCT-116/IL-18 xenograft tissuesas shown by immunohistochemistry analysis (P<0.01). Conclusion: IL-18 over-expression inhibited the growth and proliferation of HCT-116 cells both in vitro and in vivo, and the mechanism might berelated with IL-18 regulating cell cycle and promoting DNA damage.

Effects of PRDX1 gene silencing on invasion and migration of human colorectal cancer SW480 cells / 中国免疫学杂志

Objective:To investigate the effects of RNA interference(RNAi)-mediated silencing of Peroxiredoxin 1(PRDX1)gene on the invasion and migration of human colorectal cancer SW480 cells.Methods: Lentiviruses negative control vector and PRDX1 RNAi were transfected respectively into colorectal cancer SW480 cells.The transfected cells were divided into PRDX1 silencing group(si-PRDX1)and negative control group(Vector).The expressions of PRDX1 mRNA and protein in SW480 cells were exa mined by quantitative real-time PCR(qRT-PCR)and immunoblotting(Western blot),respectively.The cell migration and invasion capabilities were evaluated with transwell chamber assay and transwell chamber,respectively.The protein expressions of TIMP-2,MMP-2 and MMP-9 were detected by Western blot.Results: Compared with control group,the expressions of PRDX1 mRNA and protein were significantly decreased in PRDX1 silencing group(P<0.01),PRDX1 gene silencing cell line was successfully constructed.The levels of invasion and migration capacities of SW480 cells transfected with si-PRDX1 were lower than those in the cells transfected with control-siRNA(vector)(P<0.01).The expression of TIMP-2 was significantly increased,while the expressions of MMP-2 and MMP-9 were significantly decreased(P<0.05).Conclusion: Silencing of PRDX1 inhibits the invasion,migration and metastasis of human colorectal cancer SW480 cells by regulating the expressions of TIMP-2,MMP-2 and MMP-9.

Construction of Peroxiredoxin 2 gene lentiviral expression vector and effect on colorectal cancer SW480 cells / 中国免疫学杂志

To construct the lentiviral vector containing Peroxiredoxin 2(Prdx2) gene and the colorectal cancer cell line stably transduced with Prdx 2-containing vector , so as to provide a useful tool for studying the role of Prdx 2 in colorectal cancer.Methods: Prdx2 was amplified by PCR and inserted into lentiviral expression vector Ubi-MCS-EGFP-IRES-Puromycin (GV218) to generate Ubi-Prdx2-EGFP-Puromycin(LV-Prdx2) vector.The inserted Prdx2 gene was verified by double enzyme digestion and DNA sequencing.Subsequently ,lentiviruses were produced and transduced into SW 480 cells.EGFP expression was examined under fluorescence microscopy ,the expression of Prdx2 was detected with qRT-PCR and Western blot.Cell growth and colony forming ability were detected with MTT and plate cloning technique.Results: The lentiviral Prdx2 expression vector was successful construc-ted.Overexpression of Prdx2 was verified in SW480 cells with LV-Prdx2 vector.Prdx2 promoted SW480 cell growth and colony forming ability(P<0.05).Conclusion:Ubi-Prdx2-EGFP-Puromycin(LV-Prdx2) vector is successfully constructed,and the SW480/LV-Prdx2 cell line with stable transduction of Prdx2 containing vector is established.Overexpression Prdx2 can significantly promote the proliferation of colorectal cancer SW 480 cells.

APRIL mediates migration and invasion of colorectal cancer cells via regulation of matrix metalloproteinases / 中华微生物学和免疫学杂志

Objective To investigate the effects of a proliferation-inducing ligand(APRIL) on migration and invasion of colorectal cancer (CRC) and matrix metalloproteinases (MMPs) expression in order to observe the role of APRIL in CRC metastasis.Methods The siRNA plasmid vector targeting APRIL gene (siRNA-APRIL) was transfected into SW480 cells and recombinant human APRIL(rhAPRIL) was used to stimulate HCT-116 cells.Tumor cell migration and invasion were measured by Transwell chambers.RT-PCR and ELISA were applied to examine the expression level of MMPs.Results Metastatic and invasive capacities of siRNA-APRIL transfected SW480 were significantly inhibited,and these capacities of APRIL stimulated HCT-116 cells were significantly enhanced compared with their respective controls( all P＜0.05 ),accompanied with the alterations of MMPs mRNA and secreted protein expression( P＜0.05).The number of invading cells of SW480 control and rhAPRIL stimulated HCT-116 was significantly decreased by a MMP inhibitor GM6001 ( P＜0.05 ).Conclusion APRIL facilitates migration and invasion of CRC via regulation of MMPs,which suggests that APRIL might be used as a new target for the intervention and treatment of CRC metastasis.