Effect of circLPAR3 on radiosensitivity of esophageal cancer cells by targeting miR-1238 / 中华放射肿瘤学杂志

Objective:To evaluate the effect of circLPAR3 on the radiosensitivity of esophageal cancer cells and investigate its mechanism.Methods:The cancer tissues and and adjacent tissues of 37 patients with esophageal cancer were collected, and esophageal cancer cell lines Eca-109, EC9706 and KYSE30 and esophageal epithelial cells HET-1A were cultured in vitro. The expression levels of circLPAR3 and miR-1238 in the tissues and cells were measured by RT-qPCR. Eca-109 cells were transfected with circLPAR3 siRNA and miR-1238 mimics or co-transfected with circLPAR3 siRNA and miR-1238 inhibitor. Cell cloning experiment was conducted to evaluate the effects of silencing circLPAR3, overexpressing miR-1238, or silencing both circLPAR3 and miR-1238 on the radiosensitivity of Eca-109 cells. After Eca-109 cells that silenced circLPAR3, overexpressed miR-1238 or silenced both circLPAR3 and miR-1238 were exposed to 4 Gy irradiation, CCK-8 assay (A value), flow cytometry and Western blot were employed to assess the effects of silencing circLPAR3, overexpressing miR-1238, or silencing both circLPAR3 and miR-1238 combined with 4 Gy irradiation on the proliferation and apoptosis of Eca-109 cells and the expression levels of CyclinD1, p21, Bcl-2 and Bax proteins. Dual luciferase reporter gene experiment and RNA pull down experiment were performed to verify the regulatory relationship between circLPAR3 and miR-1238. Results:Compared with adjacent tissues, the expression level of circLPAR3 was up-regulated in the esophageal cancer tissues ( P<0.05), while that of miR-1238 was down-regulated ( P<0.05). Compared with HET-1A cells, the expression levels of circLPAR3 were up-regulated in the esophageal cancer cell lines Eca-109, EC9706 and KYSE30(all P<0.05), whereas those of miR-1238 were down-regulated (all P<0.05). Silencing circLPAR3 or overexpressing miR-1238 reduced the survival fraction of Eca-109 cells (all P<0.05), and the sensitization ratio was 1.21 and 1.75, respectively. Silencing circLPAR3 or overexpressing miR-1238 decreased the A value of Eca-109 cells and the expression levels of CyclinD1 and Bcl-2 proteins (all P<0.05), while increased the apoptosis rate of Eca-109 cells and the expression levels of p21 and Bax proteins (all P<0.05). After silencing circLPAR3 or overexpressing miR-1238 combined with 4 Gy irradiation, the A value of Eca-109 cells and the expression levels of CyclinD1 and Bcl-2 proteins were decreased (all P<0.05), while Eca-109 cell apoptosis rate and the expression levels of p21 and Bax proteins were increased (all P<0.05). circLPAR3 targeted and negatively regulated the expression level of miR-1238 in Eca-109 cells. After silencing miR-1238 and circLPAR3 simultaneously, the survival fraction of Eca-109 cells was higher than that when only silencing circLPAR3, and the sensitization ratio was 0.59. Silencing miR-1238 reversed the effects of silencing circLPAR3 combined with 4 Gy irradiation on the proliferation and apoptosis of Eca-109 cells. Conclusion:circLPAR3 is highly expressed in esophageal cancer tissues and cell lines, and silencing the expression of circLPAR3 can inhibit the proliferation of esophageal cancer Eca-109 cells, promote their apoptosis, and enhance cell radiosensitivity by up-regulating miR-1238.

Clinical Significance of Detection of ZIC1 Promoter Methylation in Peripheral Blood and Cancer Tissues of Non-small Cell Lung Cancer Patients / 肿瘤防治研究

Objective To investigate the methylation status of ZIC1 gene in peripheral blood and lung cancer tissues of NSCLC patients and its prognostic significance. Methods We took the peripheral blood, cancer tissues and adjacent tissues of 95 NSCLC patients. The peripheral blood of 95 healthy people was taken as control group. MSP was used to compare the detection rate of ZIC1 methylation between peripheral blood and cancer tissues. And we analyzed the correlation of ZIC1 methylation in peripheral blood and cancer tissues with the clinicopathological factors of NSCLC patients. Results The methylation detection rates of ZIC1 in peripheral blood and lung cancer tissues in NSCLC patients were significantly higher than those in peripheral blood of healthy people and adjacent tissues of NSCLC patients (P < 0.05), and the sensitivity and specificity of ZIC1 methylation in the diagnosis of tumor tissues were higher. The positive rate of ZIC1 gene methylation in peripheral blood and tumor tissues of NSCLC patients was significantly correlated with tumor diameter, metastasis, stage and pleural effusion (P < 0.05). Conclusion The methylation of ZIC1 gene is related to the occurrence, development, metastasis and stage of NSCLC. It may be used as a diagnostic and prognostic indicator of NSCLC.

Expression of tRF 31 in breast cancer tissues and its effect on cancer cell proliferation / 医学研究生学报

ObjectiveAt present, there are few reports on the effect of tRF 31 on breast cancer. This study aims to detect and analyze the expression level of tRF 31 in breast cancer tissues and to explore its effect on the proliferation of breast cancer cells.Methods32 tumor tissue samples and the corresponding para-cancer tissues from breast cancer patients in The Affiliated Cancer Hospital of Nanjing Medical University from November 2017 to February 2019 were collected. Six out of the thirty two samples were selected for high-throughput sequencing and at last tRF 31 (FC=6.5781, P=0.023) was selected as the research object. The expression of tRF-31 in cancer and para-cancer tissues was measured by RT-PCR. The sensitivity and specificity of tRF-31 expression in the breast cancer diagnosis were analyzed by ROC curve. Two kinds of human breast cancer cell lines (MDA-MB-231 and MCF-7) were divided into two groups: control group (transfected with negative control) and tRF-31 group (transfected with tRF-31 inhibitor). The proliferation of transfected breast cancer cells was detected by CCK-8 and clonal formation assay. The expression changes of threonine kinase (AKT) and mammalian target of rapamycin (mTOR) in breast cancer cells after transfection were measured to explore the relationship between TRF-31 and AKT/mTOR signaling pathway.ResultsThe expression of tRF-31 in cancer tissues (0.103±0.207) was significantly higher than that in para-cancerous ones (0.028±0.039). ROC curve showed that the sensitivity and specificity of tRF-31 in detecting breast cancer were 90.63% and 53.13%, respectively. In MDA MB 231 cells, the expression of tRF-31 in the tRF-31 group was significantly lower than that in the control group [(0.267±0.012) vs (1±0.040), P<0.01)], and in MCF-7 cells, the expression of tRF-31 in the tRF-31 group was also significantly lower than that in the control group. In MDA-MB-231 and MCF-7 cells, the proliferation ability of the tRF-31 group was lower than that of the control group at 0h, 24h, 48h and 72h. In MDA-MB-231 cells, the clonal formation rate of tRF-31 group [(43.67±3.29)%] was significantly lower than that of the control group [(100±3.74)%] (P<0.01). In MCF-7 cells, the clonal formation rate of tRF-31 group [(49±2.94)%] was significantly lower than that of the control group [(100±4.89)%] (P<0.01). In MDA-MB-231 and MCF-7 cells, the protein levels of AKT and mTOR in tRF-31 group were significantly lower than those in the control group.ConclusiontRF-31 is highly expressed in breast cancer tissues and can promote the proliferation of breast cancer cells. This result is expected to provide a new target for the treatment of breast cancer.

Application of lncRNA AK057037 in tissues and blood samples of patient with hepatocellular cancer / 国际检验医学杂志

Objective To investigate the clinical value of tissue and blood long noncoding RNA(lncRNA) detection in the patients with hepatocellular cancer(HCC).Methods The HCC tissue sample,blood sample, suspected HCC puncture sample and blood sample from the persons undergoing healthy physical examination were selected for conducting the study.The six lncRNAs which were more than 5-fold expression amount were picked out in the 338 transcripts with abnormal high expression.The expression amount of lncRNA AK057037 in HCC tissue and serm tissue was detected by using the real time quantitative polymerase chain reaction and in situ hybridization.Then the results were statistically analyzed.Results The six lncRNAs were highly ex-pressed in the tissues and blood of the liver cancer,and the difference was statistically significant(P<0.05). Compared with healthy people,the relative expression of AK057037 in the tissues and blood of the liver cancer increased significantly,and the relative expression level in tumor and paracancerous tissues was statistically significant(P<0.05).As the tumor progresses,the relative expression of AK057037 was further enhanced. There was a higher relative expression in advanced liver cancer especially,and the difference was statistically significant(P<0.05).The results of the receiver operating characteristic curve showed that the area under curve of AK057037 was 0.834(95% CI 0.705-0.918)in the blood specimen,and its diagnostic efficiency was better than that of commonly used nucleic acid marker miR-29a.Conclusion lncRNA AK057037 is highly ex-pressed in HCC sample and blood,and can be used for clinical detection of HCC.

In Vivo Observation of Endothelial Cell-Assisted Vascularization in Pancreatic Cancer Xenograft Engineering

In this study, for better understanding of patient-derived xenograft (PDX) generation, angiogenic characteristics during PDX cancerous tissue generation was investigated with different initial cell seeding conditions in the hydrogel. We monitored the angiogenic changes during the formation of in vivo cancer cell line xenografts induced by endothelial cells. Our in vivo cancer tissue formation system was designed with the assistance of tissue engineering technology to mimic patient-derived xenograft formation. Endothelial cells and MIA PaCa-2 pancreatic carcinoma cells were encapsulated in fibrin gel at different mixing configurations and subcutaneously implanted into nude mice. To investigate the effect of the initial cancerous cell distribution in the fibrin gel, MIA PaCa-2 cells were encapsulated as a homogeneous cell distribution or as a cell aggregate, with endothelial cells homogeneously distributed in the fibrin gel. Histological observation of the explanted tissues after different implantation periods revealed three different stages: isolated vascular tubes, leaky blood vessels, and mature cancerous tissue formation. The in vivo engineered cancerous tissues had leaky blood vessels with low expression of the vascular tight junction marker CD31. Under our experimental conditions, complex cancer-like tissue formation was most successful when tumorous cells and endothelial cells were homogeneously mixed in the fibrin gel. The present study implies that tumorous xenograft tissue formation can be achieved with a low number of initial cells and that effective vascularization conditions can be attained with a limited volume of patient-derived cancer tissue. Endothelial cell-assisted vascularization can be a potent choice for the effective development of vascularized cancerous tissues for studying patient-derived xenografts, cancer angiogenesis, cancer metastasis, and anticancer drugs.

Mutation analysis of autosomal and X chromosomal STR in gynecologic and breast cancer / 中国法医学杂志

Objective To explore the mutation types and disciplines of STR commonly used in forensic in gynecologic and breast cancerand investigate the application of microdissection in forensic practice involving tumor tissue. Methods DNA of tumor tissues, adjacent normal tissues and peripheral blood from 62 patients with breast cancer, 62 patients with gynecologic cancer and 10 patients with benign gynecologic tumor were amplified by PowerPlex 21 System kit and Argus X-12 kit. Capillary electrophoresis of PCR products was carried out on an ABI 3130 Genetic Analyzer to obtain genotypes. Some tumor tissues with STR variation were microdissected. Results The genotype of peripheral blood in cancer patient was consistent with that of corresponding normal tissue. 4 types of STR variations were found in 46.77% gynecologic cancer tissues, compared with that in benign tumor tissues and breast cancer, the difference of STR variation was significant(P<0.01,P=0.009). The genotype of stromal cells separated by microdissection was consistent with that of corresponding adjacent normal tissue. Conclusion The STR loci detected in the study with poor stability are not suitable for forensic cases involving gynecologic cancer tissues. The genotype of stromal cells separated accurately from tumor tissues by microdissection could represent the normal DNA genotype of the individual with cancer. Microdissection is an effective solution in forensic cases with tumor tissues.

The expression of microRNA-21,microRNA-135b and microRNA-141 in colon cancer tissue and their correlation / 国际检验医学杂志

Objective To investigate the expression level of microRNA-21,microRNA-135b and microRNA-141 in colon cancer tissue.Methods Totally 40 cases of colon cancer specimen from surgical resection in our hospital from May 2013 to April 2014 were selected,40 healthy colon tissues from people who entered our hospital at the same period for physical examination were selected.The expression level of microRNA-21,microRNA-135b and microRNA-141 were determined using real time PCR.Chosed sex,age,lymph node metastasis,TNM staging,differentiation degree as related factors by literatures published in home and abroad,assessed the correlation between the level of microRNA-141,microRNA-135b,microRNA-21 and its expression in colorectal cancer.Results The expression level of microRNA-21,microRNA-135b and microRNA-141 in colon cancer tissues were remarkably higher than those in healthy tissues,the difference was statistically significant(P<0.05).The expression levels of microRNA-141,microRNA-135b and microRNA-21 in colon cancer tissues were closely related to the occurrence of lymph node metastasis,progression and stage of colon cancer,and the difference was statistically significant(P<0.05).Conclusion The expression of microRNA-21,microRNA-135b and microRNA-141 are upregulated in colon cancer tissue,which is highly correlated with the lymphatic metastasis and progressiveness of colon cancer.

Expression of DEK Protein in the Tissue and Voided Urine of Patients with Bladder Urothelial Carcinoma and Its Clinical Significance / 中国医科大学学报

Objective To detect the presence of DEK protein in the tissue and voided urine of patients with bladder urothelial carcinoma ,and an?alyze the relationship between DEK protein and clinical pathological classification of bladder cancer. Methods The expression of DEK protein was examined by immunohistochemistry in 48 bladder cancer tissues and 18 adjacent normal tissues(control group). The presence of DEK protein in voided urine was analyzed by western blot in 28 bladder cancer patients and 6 healthy individuals(control group). Results The positive ex?pression of DEK protein in bladder tumor tissues(73%)is higher than in adjacent normal tissues(33%,P<0.05). The expression of DEK in su?perficial bladder cancer(86%)was found higher than invasive bladder cancer(55%)with significant differences(P<0.05). The expression of DEK in urine samples of bladder cancer patients(0.834)was found higher than control group(0.242,P<0.05);Compared with the DEK expres?sion of control group,the sensitivity of diagnosis of bladder cancer with voided urine could be 82.1%. Conclusion DEK protein positive ex?pressed in tissues and voided urine of patients with bladder urothelial carcinoma. The expression was correlated with pathological classification of bladder cancer. The positive rate of DEK expression in early stage of tumor tissues is higher than late stage. The presence of DEK protein in tissues and voided urine can be considered as a suitable biomarker for bladder cancer potentially.

Correlation between Expression of MMP-9, TIMP-1 and VEGF and the Invasion of Non Small Cell Lung Cancer / 昆明医科大学学报

Objective To investigate the relationship between metalloproteinase-9-9 (Matrix),inhibitor-1-1 (MMP-9),metalloproteinase TIMP-1 factor (Matrix) and endothelial growth (VEGF),and to investigate the correlation between the expression of (matrix) and vascular endothelial growth factor (endothelial) and vascular endothelial growth factor (vascular).Methods Sixty-eight cases of non small cell lung cancer patients treated in fengning manchu autonomous county hospital from January to January 2014 were analyzed retrospectively,and the expression level of TIMP-1,VEGF and MMP-9 were analyzed by immunohistochemical method.Results The positive rates of MMP-9 (75％),TIMP-1 (19.12％) and VEGF (13.24％) were significantly higher than those in normal tissues (P＜0.05),MMP-9 (60.87％),VEGF (54.55％),TIMP-1 (65.22％) and VEGF (76.09％,59.09％,63.64％).The positive rates ofTIMP-1,VEGF and MMP-9 were higher than those without lymph node metastasis (P ＜0.05) Conclusion MMP-9,TIMP-1 and VEGF expression levels can predict the invasion and metastasis of non small cell lung cancer,and the three kinds of substances can promote the development and progression of lung cancer in different degrees.

Expression and clinical significance of NrF-2 and Ho-1 in gastric carcinoma / 中国生化药物杂志

Objective To study nuclear factor E-2 related factor 2(NrF-2) and heme oxygenase -1(Ho-1) expression and to explore its clinical significance.Methods From February 2016 to June 2016, 176 cases human gastric tissue paraffin blocks selected in department of pathology in Hangzhou Cancer Hospital, including 78 cases of gastric cancer, 78 cases of normal tissue were analyzed.And the expressions of NrF-2 and Ho-1 were detected by immunohistochemistry in two groups of human gastric tissue.Results The levels of NrF-2 and Ho-1 in gastric cancer group were(0.752 ± 0.098),(0.862 ±0.081) were significantly higher than those in normal group(0.381 ±0.068),(0.412 ±0.083);NrF-2, the positive expression rate of Ho-1 in gastric wall invasion degree of T3 +T4 with lymph node metastasis, NTM stage III and IV gastric cancer tissues were significantly increased.The differences were statistically significant ( P <0.05 ) .Conclusion NrF-2 and Ho-1 play the important role in the development of gastric cancer and NrF-2, the positive expression rate of Ho-1 in the gastric carcinoma and gastric wall invasion, lymph node metastasis and NTM staging are closely related.The clinical diagnosis if the combined detection of NrF-2 and Ho-1, is conducive to the early diagnosis of gastric cancer and the prognosis of the disease.

Free amino acids differential analysis between breast cancer and mammary glandular tissues / 中华内分泌外科杂志

Objective To assay free amino acids (FAA) content of breast cancer tissue and mammary glandular tissue and discuss the therapeutic basis of unbalanced amino acids of breast cancer. Methods The selected samples were conserved and then cut into pieces, grinded, syruped and deproteinized. Then FAA were separated from the samples. The content of 17 kinds of free amino acids in breast cancer tissue and mammary glandular were assayed respectively. Results The concentration of FAA in breast cancer tissue was higher than that in mammary glandular tissue. Conclusions Free amino acids metabolism was greatly different in breast cancer and mammary glandular tissues. This offers a therapeutic basis for the treatment of unbalanced free amino acids in the case of breast cancer.

Enhancement of Y14 and Upf1 expression in human breast cancer cell lines / 基础医学与临床

Objective To study expression enhancement and significance of Y14 and Upf1 in human breast cancer cell lines and tissue. Methods Immuocytochemistry and laser scanning confocal microscope(LSCM) were applied. Y14 and Upf1 were determined in human breast cancer cell lines(MCF-7,ZR-75-30,T47D,MDA-MB435s,MDA-MB-453, MDA-MB-231) and breast epithelial cell line ( HBL-100). Results (1) Y14 and Upf1 level of breast cancer cells are obviously higher than that in breast epithelial cell line (P < 0. 05 ). (2)Y14 and Upf1 level of MDA-MB-231 are obviously higher than that in MCF-7. (3)The expression enhancement of Y14 and Upf1 level are obviously higher in human breast cancer tissue. Conclusion The expression level of Y14 and Upf1 in breast cancer cells and tissue enhance obviously.

Expression and Significance of Apoptosis Related Gene BAG-1 in Digestive Tract Cancer by Tissue Microarray / 中国康复理论与实践

@#ObjectiveTo detect BAG1 expressions in digestive tract cancer by tissue microarray and to evaluate its clinical significance.MethodsTissue microarray of digestive tract cancer and normal tissues were analyzed by DAKO Envision system immunohistochemical staining for apoptosis related gene BAG-1 expression.ResultsThe positive rate of BAG-1 expression among esophagus cancer,gastric cancer and rectal cancer were higher than that of normal tissues respectively(P<0.01).ConclusionThere is an overexpression of BAG-1 in digestive tract cancer,which suggest that apoptosis related gene BAG-1 may be related to these cancer.

Enhancement of Y14 and Upf1 expression in human breast cancer cell lines / 基础医学与临床

Objective To study expression enhancement and significance of Y14 and Upf1 in human breast cancer cell lines and tissue.Methods Immuocytochemistry and laser scanning confocal microscope(LSCM) were applied. Y14 and Upf1 were determined in human breast cancer cell lines(MCF-7,ZR-75-30,T47D,MDA-MB-435s,MDA-MB-453,MDA-MB-231)and breast epithelial cell line(HBL-100).Results (1)Y14 and Upf1 level of breast cancer cells are obviously higher than that in breast epithelial cell line(P

Application of laser capture microdissection to capture pure cells in lung cancer and their paired normal lung tissues / 西安交通大学学报(医学版)

Objective Microheterogeneity is an important problem in molecularly biological and proteomic research on malignancies.The goal of this study was to investigate the method to purify targeted cells in lung cancer and its paired normal tissues for proteomic study on lung cancer applying laser capture microdissection(LCM).Methods LCM technique was emploged to obtain the cells of lung cancer tissues and their paired normal tissues from frozen sections.Results LCM can be applied to quickly and precisely obtain pure targeted cells subgroup or single cell without interstitium under the microscope.Conclusion LCM can tackle the problem of tissue heterogeneity in molecular analysis successfully.These results indicate that LCM has a potential as a tool in lung cancer proteomic research.

Detecting the levels of serum TPS、CYFRA21-1 and STNFR in patients with lung cancer and its clinical value / 中国实用内科杂志

Objective To investigate the diagnostic value of tissue polypeptide specific antigen (TPS),CYFRA21-1 and soluble tumor necrosis factor receptor (STNFR) in patients with lung cancer.Methods The serum levels of TPS,CYFRA21-1 and STNFR were determined by enzyme-linked immunosorbent assay (ELISA) in 72 patients with lung cancer,54 patients with benign diseases and 32 healthy adults.Results The levels of the 3 tumor markers in lung cancer group were significantly higher than those of the benign disease group and healthy control group,and the serum levels of this 3 tumor markers corresponding increased with the high TNM stage.The detectable rates of TPS (80.5%) and STNFR (81.9%) in serum of lung cancer were higher than CYFRA21-1(65.3%),and in TNM I group,the detectable rates of TPS (57.1%) and STNFR (57.2%) were higher than CYFRA21-1 (25.6%) too.Conclusion The TPS,CYFRA21-1 and STNFR can be used as a very useful and sensitive tumor marker in the diagnosis of lung cancer.The TPS and STNFR are better than CYFRA21-1 in clinical use.

Comparison of c-erbB-2 Oncoprotein Expression in Serum and Tissue of Gastric Cancer Patients / 대한임상병리학회지

BACKGROUND: A soluble fragment of the c-erbB-2 oncoprotein become detectable in the serum of the gastric cancer patients by enzyme-linked immunosorbent assay(ELISA). METHODS: Serum c-erbB-2 levels of 67 gastric cancer patients are measured. In 27 patients, tissue levels of c-erbB-2 in paired normal and gastric cancer tissues are measured by ELISA(Triton Diagnostics Inc., USA). RESULTS: As 14 U/mL for positivity, the sero-positivity of the c-erbB-2 was 33%(22/67). The positivity showed no differences in cancer stages and tumor differentiations(well-moderate 29%, poor-signet ring cell type 35%). Tissue c-erbB-2 level was 390+/-284 U/mL and 568+/-536 U/mL in normal and cancer tissue, respectively. Blood c-erbB-2 level did not correlate with either cancer tissue c-erbB-2 level or normal tissue c-erbB-2 level. Nineteen out of 27 patients showed higher c-erbB-2 levels in cancer than normal tissues. The positivity of c-erbB-2 in serum was 7 out of 19 c-erbB-2 positive cancer tissue, whereas the positivity in serum was one out of 8 c-erbB-2 negative cancer tissue. CONCLUSIONS: Elevated blood c-erbB-2 oncoprotein level might reflect elevated c-erbB-2 level of gastric cancer tissue.