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OBJECTIVES@#The high morbidity and mortality of colorectal cancer (CRC) have posed great threats to human health. Circular RNA (circRNA) and microRNA (miRNA), acting as competing endogenous RNAs (ceRNAs), have been found to play vital roles in carcinogenesis. This paper aims to construct a circRNA/miRNA/mRNA regulatory network so as to explore the molecular mechanism of CRC.@*METHODS@#The sequencing data of circRNA from CRC were obtained from Gene Expression Omnibus (GEO). The differential circRNA was screened and its structure was identified by Cancer-specific CircRNA Database (CSCD); the sequencing data of miRNA and messenger RNA (mRNAs) were downloaded from The Cancer Genome Atlas (TCGA) database and the differentially expressed genes were screened; the corresponding miRNA of differential circRNAs were predicted by CircInteractome database; DIANA, Miranda, PicTar, and TargetScan databases were used to predict the target genes of different miRNAs; the target genes from Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were enriched by R language; String database combined with Cytoscape 3.7.2 software was used to construct protein-protein interaction (PPI) network and hub genes were screened; the expressions of mRNAs in the Top10 hub genes were verified in CRC. The network diagrams of circRNAs/miRNAs/mRNAs and circRNAs/miRNAs/Top10 hub mRNAs were constructed by Cytoscape3.7.2. Real-time PCR was used to examine the expression levels of hsa_circRNA_0065173, hsa-mir-450b, hsa-mir-582, adenylate cyclase 5 (ADCY5), muscarinic acetylcholine receptor M2 (CHRM2), cannabinoid receptor 1 (CNR1), and lysophosphatidic acid receptor 1 (LPAR1) in the CRC tissues and the adjacent normal tissues.@*RESULTS@#A total of 14 differential circRNAs were identified, and 8 were found in CSCD; 34 miRNAs targeted by circRNAs were obtained. The PPI network was constructed, and the Top10 hub genes were identified, which were CHRM2, melanin concentrating hormone receptor 2 (MCHR2), G-protein gamma 3 subunit (GNG3), neuropeptide Y receptor Y1 (NPY1R), CNR1, LPAR1, ADCY5, adenylate cyclase 2 (ADCY2), gamma 7 (GNG7) and chemokine 12 (CXCL12), respectively. The expressions of Top 10 hub genes were also verified, and the results showed that the Top 10 hub genes were down-regulated in CRC; the constructed network diagram showed that hsa_circRNA_0065173 may regulate ADCY5, CHRM2, and Hsa-mir-450b by modulating hsa-mir-450b and hsa-mir-582. CNR1 and LPAR1 genes might serve as potentially relevant targets for the treatment of CRC. Real-time PCR results showed that the expression levels of hsa_circRNA_0065173, ADCY5, CHRM2, CNR1 and LPAR1 in the CRC tissues were significantly reduced compared with the adjacent normal tissues (all P<0.05); the expression levels of hsa-mir-450b and hsa-miR-582 were significantly increased (both P<0.05).@*CONCLUSIONS@#In this study, a potential circRNAs/miRNAs/mRNAs network is successfully constructed, which provides a new insight for CRC development mechanism through ceRNA mediated by circRNAs.
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Humans , Colorectal Neoplasms/genetics , Computational Biology/methods , Gene Regulatory Networks , MicroRNAs/genetics , RNA, Circular/genetics , RNA, Messenger/geneticsABSTRACT
Objective:To investigate the prognostic value of systemic inflammatory score (SIS) in prognostic assessment of patients with unresectable metastatic colorectal cancer (mCRC).Methods:The clinical data of 130 patients with unresectable mCRC in Affiliated Haikou Hospital of Xiangya Medical College of Central South University from January 2014 to December 2016 were retrospectively analyzed. The relationship between SIS and clinicopathological characteristics of unresectable mCRC patients was also analyzed. The survival analysis was made by using Kaplan-Meier. The risk factors affecting the prognosis of unresectable mCRC patients were analyzed by using Cox regression model to make univariate and multivariate analysis.Results:According to SIS results, patients were divided into 0-score group (40 cases), 1-score group (58 cases), and 2-score group (32 cases). There were no significant differences in different SIS constitution patients stratified by age, gender, primary tumor location, functional status score, tissue type, RAS gene status, number of metastatic organs, peritoneal spread and molecular targeted therapy (all P > 0.05). Kaplan-Meier survival analysis showed that the 5-year overall survival rates of SIS 0-score, 1-score and 2-score group were 37.5%, 19.0%, 6.3%, respectively; and the difference was statistically significant ( χ2 = 3.152, P<0.01). Cox regression survival analysis showed that female, primary tumor location in right side and SIS (scores of 1-2) were independent risk factors for overall survival in patients with unresectable mCRC (all P<0.05). Conclusion:SIS may be an important indicator for prognostic assessment of patients with unresectable mCRC, and patients with high SIS have poor prognosis.
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Objective:To explore the clinical significance of classification and grading of ovarian serous tumors (OST) and the correlation of classification and grading with expressions of p53 and Ki-67 proteins, so as to provide a basis for accurate diagnosis and reasonable treatment of OST.Methods:A total of 100 paraffin-embedded ovarian tissues were collected from Haikou Affiliated Hospital of Central South University Xiangya School of Medicine from January 2012 to November 2017. The classification and grading of OST was based on the World Health Organization (WHO) ovarian tumor tissue classification, the work conference on borderline ovarian tumors which was held in August 2013 in Maryland, United States, and the two-level tissue classification system of the United States MD Anderson Cancer Center. The 100 cases of ovarian tissues included 10 cases of normal ovarian tissues (NOT), 12 cases of ovarian serous cystadenoma (OSA), 18 cases of ovarian serous borderline tumor (OSBT), 22 cases of low-grade serous carcinoma (LGSC), and 38 cases of high-grade serous carcinoma (HGSC). The expressions of p53 and Ki-67 proteins in paraffin-embedded tissues were detected by immunohistochemistry. The clinical characteristics of patients with different types of OST and their relationships with the expression of p53 protein and Ki-67 positive index were analyzed.Results:Among OSBT, LGSC and HGSC groups, the proportion of patients with onset age > 50 years old [38.9% (7/18), 45.5% (10/22), 73.7% (28/38)], poor differentiation [0 (0/18), 0 (0/22), 100.0% (38/38)], stage Ⅲ-Ⅳ [5.6% (1/18), 27.3% (6/22), 39.5% (15/38)] increased sequentially, and the differences were statistically significant (all P < 0.05). Among NOT, OSA, OSBT, LGSC and HGSC groups, there were significant differences in the positive rate of p53 protein [20.0% (2/10), 25.0% (3/12), 27.8% (5/18), 31.8% (7/22), 57.9% (22/38)] and Ki-67 positive index [(10.40±0.00)%, (31.49±6.53)%, (42.81±6.84)%, (74.29±6.54)%, (77.04±8.88)%] (all P < 0.05). In the patients with ovarian serous carcinoma (LGSC + HGSC), there was no significant difference in the positive rate of p53 protein and Ki-67 positive index between the onset age ≤ 50 years old and > 50 years old groups (both P > 0.05), but there were significant differences between the patients with different tissue differentiation grade, clinical stage and metastasis (all P < 0.05). There was significant difference in Ki-67 positive index among NOT, OSA, OSBT, LGSC and HGSC patients with positive expression of p53 protein ( P < 0.01). Conclusions:The classification and grading of OST is related to onset age, tissue differentiation and clinical stage, which can be used to guide treatment and judge prognosis. The positive rate of p53 protein and Ki-67 positive index increase with advancing grade of breast lesions, which may be related to the occurrence and development of OST.
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Objective To investigate the role of PKCι, YAP1 and high-risk HPV infection in the local immune microenvironment of cervical cancer. Methods We chose 80 cases of normal tissue of the cervix (NCT), cervical low-grade squamous intraepithelial lesion (LSIL), cervical high-grade squamous intraepithelial lesion (HSIL) and early cervical squamous cell carcinoma (SCC) each. Four groups were collected.The infection rate of high-risk HPV in four groups was determined by real-time fluorescence PCR method. The expression levels of PKCι, YAP1, CD4 and CD8 in four groups were measured and correlated by IHC and clinicopathologic features were also analyzed. Results The differences of high-risk HPV infection rate and PKCι, YAP1, CD4, CD8 positive rate among groups of NCT, LSIL, HSIL and SCC had statistical significance (P < 0.05). The level of cervical lesions was positively associated with high-risk HPV infection and positive PKCι, YAP1, CD8 expression (P < 0.05), while negatively associated with positive CD4 expression (P < 0.05). HPV infection and positive PKCι, YAP1, CD8 expression were positively correlated with each other in SCC, while were all negatively correlated with positive CD4 expression(P < 0.05). The differences of HPV infection, PKCι, YAP1 and CD8 positive expression were significant in different levels of differentiation and vascular invasion of SCC (P < 0.05). Conclusion The patients with cervical lesions are often accompanied by high-risk HPV infection and abnormal expression of PKCι, YAP1, CD4 and CD8, which may have synergistic effects on each other, causing the local immunosuppression microenvironment of SCC. It provides a possible strategy for the study of pathogenesis, diagnosis and treatment of cervical cancer.
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Objective To study the specific killing effect in human carcinoma cells aftercombination treatment of radiation and p53 gene regulated by a radiation-enhanced promoter.Methods Aplasmid pE6 (TATA)-p53 was constructed.After irradiation,the expression of P53 was detected withWestern blot assay,apoptosis was detected by Annexin V-FITC,and cell survival was detected byclonogenic assay then the sensitivity enhancement ratio (SER) was analyzed for HeLa and A549 cells.Results The expression of P53 were increased in the irradiated cells and 6 Gy irradiation triggered thestrongest activity.After p53 transfection,radiation-induced apoptosis was obviously enhanced incomparison with the control group without gene transfection (F =11.018,10.736,P ＜ 0.05).The SER ofp53-promoter was 2.36 for A549 cells and 2.56 for Hela cells.Conclusions The p53-plasmid promotercould induce apoptosis and enhance the radiosensitivity of tumor cells,which may provide a noveltherapeutic strategy for cancer treatment.