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OBJECTIVE： To investigate the correlation of ADPRT rs1136410 polymorphism with the occurrence of non-small cell lung cancer （NSCLC） in Han nationality from northern Jiangsu. METHODS： A total of 283 patients with primary NSCLC of Han nationality in Northern Jiangsu were selected from the Affiliated Hospital of Xuzhou Medical University during Nov. 2015-Dec. 2018 as NSCLC group. A total of 210 healthy subjects underwent physical examination were included in control group. PCR-RFLP was utilized to determine the genotypes at ADPRT rs1136410 locus. Logistic regression model was used to evaluate the effect of polymorphism and its interaction with smoking on the occurrence of NSCLC. RESULTS： There was no statistical significance in age and gender between 2 groups （P＞0.05）. The proportion of smoker in NSCLC group was significantly higher than control group （P＜0.05）. TT， TC and CC genotypes were detected at rs1136410 locus of ADPRT gene. The frequency of TT， TC and CC genotype were 41.9％，44.8％ and 13.3％， and those of allele T and C were 64.3％ and 35.7％ in control group. The frequency of TT， TC and CC genotype were 21.6％， 50.2％ and 28.2％， and those of allele T and C were 46.6％ and 53.4％ in NSCLC group， respectively. The frequencies of genotypes in 2 groups were in accordance with Hardy-Weinberg equilibrium （P＞0.05）， while there was significant difference in genotype and allele frequencies between 2 groups （P＜0.05）. Compared with TT genotype， the risk of NSCLC in individuals carrying TC and CC genotypes raised by 1.179， 3.122 folds [ORTC＝2.179， 95％CI （1.435， 3.309）， P＜0.05； ORCC＝4.122，95％CI（2.401，7.075），P＜0.05]. Compared with individuals carrying TT genotype， the risk of NSCLC occurrence in non-smokers carrying TC and CC genotypes increased by 0.371， 1.328 fold [ORTC＝1.371，95％CI （0.927，3.428），P＜0.05； ORCC＝2.328，95％CI （1.249，4.622），P＜0.05]； and the risk of NSCLC occurrence in smokers carrying TC and CC genotypes increased by 0.928， 2.182 folds [ORTC＝1.928，95％CI （1.257，2.957）， P＜0.05；ORCC＝3.182，95％CI （1.760，5.754）， P＜0.05]. CONCLUSIONS： The rs1136410 locus mutant genotype of ADPRT gene is the risk factor of NSCLC in Han nationality from Northern Jiangsu， and smoking raises this risk of NSCLC occurrence in individuals with mutation genotypes of ADPRT rs1136410.
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OBJECTIVE:To study the gene mutation status of epidermal growth factor receptor(EGFR)in non-small cell lung cancer(NSCLC)patients and its relationship with clinical indexes,and to provide reference for individualized administration of EGFR-TKI in NSCLC patients. METHODS:Totally of 274 NSCLC patients from the northern of Jiangsu area were selected from our hospital during Jan. 2015-Dec. 2017. Mutation status of EGFR gene in lung tissue was determined by amplification refractory mutation system (ARMS)-TaqMan PCR assay. The relationship of EGFR gene mutation with clinical indexes as gender,age, smoking status, staging, tumor differentiation and pathological type were analyzed retrospectively. Compared with related literatures,the regional differences of EGFR gene mutation were analyzed. RESULTS:Among 274 NSCLC patients,112 patients suffered from EGFR gene mutation with total mutation rate of 40.88%. There were 50,57,3,2 cases of exon 19,exon 21,exon 20 and exon 19+21 mutation,and the types of EGFR gene mutation were delE746-A750,L858R and insH773-V774H,etc. The mutation rates of EGFR gene exon 19,exon 21 in non-smoking,early,well-differentiated and adenocarcinoma patients were 52.50%,47.24%,46.36% and 45.00%,which were significantly higher than smoking (28.57%),advanced (27.03%), poor-differentiated(31.71%)and squamous cell carcinoma(27.66%)patients,with statistical significance(P＜0.05). There was no statistical significance in mutation rates of EGFR gene exon 19 and exon 21 between male and female,≥65 year-old and ＜65 year-old patients (P＞0.05). EGFR mutation rate of NSCLC subjects from the northern of Jiangsu area was significantly higher than Shanghai area(P＜0.05);there was no statistical significance compared with Yunnan area(P＞0.05)but mutation types were different. CONCLUSIONS:There is the highest EGFR gene mutation rate in its exon 21,lesser in exon 19,rare in exon 20 and exon 19+21 among NSCLC patients from the Northern of Jiangsu area. There are obvious regional differences. The mutation rate of EGFR gene mutation exon 19 and exon 21 are associated with smoking status,staging,tumor differentiation and pathological type of NSCLC patients. The non-smoking, early stage, well-differentiated and adenocarcinoma patients are more likely to benefit from EGFR-TKI targeted therapy.
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OBJECTIVE:To investigate the correlation of XRCC1 rs25487 polymorphism with the occurrence of lung cancer. METHODS:A total of 208 patients with primary lung cancer of Han nationality in Northern Jiangsu selected from the Affiliated Hospital of Xuzhou Medical University during Sept. 2015-Jul. 2016 were included in lung cancer group. A total of 214 healthy volunteers of the hospital underwent physical examination were included in control group. PCR-RFLP was used to detect the genotypes at XRCC1 rs25487 locus,and Logistic regression model was used to evaluate the correlation of genotypes with the occurrence of lung cancer. RESULTS:There was no statistical significance in the distribution of age and gender between 2 groups (P＞0.05). The proportion of smoker in lung cancer group was significantly higher than control group,with statistical significance(P＜0.05). AA,AG and GG genotypes were detected at rs25487 locus of XRCC1 gene. The frequency of AA,AG and GG genotype were 43.5%,41.1%and 15.4% in control group and 28.8%,48.6% and 22.6% in lung cancer group,respectively. The frequencies of genotypes in 2 groups were in line with Hardy-Weinberg equilibrium(P＞0.05),but there was statistical significance in genotype distribution between 2 groups(P＜0.05). Compared with AA genotype,the risk of lung cancer in individuals carrying AG genotype increased by 2.265 fold [OR＝2.265,95%CI(1.299,3.950),P＝0.040;after corrected with gender,age and smoking history OR＝2.309,95%CI(1.274, 4.185),P＝0.006],with statistical significance. The risk of lung cancer in individuals carrying GG genotype increased by 1.310 fold [OR＝1.310,95%CI(0.771,2.228),P＝0.318;after corrected OR＝1.429,95%CI(0.811,2.518),P＝0.217],without statistical significance. CONCLUSIONS:rs25487 locus mutant heterozy-gosity of XRCC1 gene is risk factor of lung cancer in Han nationality from Northern Jiangsu,and smoking can increase the risk of lung cancer.
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Objective: To explore the efficacy and safety of thalidomide for the treatment of delayed vomiting, induced by chemotherapy in cancer patients
Study Design: Randomized, double-blind controlled study
Place and Duration of Study: The Oncology Department of Affiliated Hospital of Xuzhou Medical University, Jiangsu Xuzhou, China, from January 2012 to January 2014
Methodology: A total of 78 cancer patients, who had delayed vomiting observed from 24 hours to 1 week after chemotherapy, were included in the study. Patients were divided in a treatment group [40 patients, 51.28%] and a control group [38 patients, 48.71%]. The treatment group received thalidomide at an oral dose of 100 mg per night; 50 mg was added daily up to a dose of 200 mg per night, if the curative effect was suboptimal and the medicine was tolerated. Both the treatment and the control groups received a drip of 10 mg azasetron 30 minutes before chemotherapy. The control group only proportions of antiemetic effects and adverse reactions were compared using the c2 test. Antiemetic effects and adverse reactions were assessed from Odds Ratios [OR] with 95% Confidence Intervals [95% CI]
Results: The effective control rate of delayed vomiting in the treatment group was significantly higher than that in the control group [c2=5.174, p=0.023]. No significant difference was found between the two groups in other adverse effects of chemotherapy. Karnofsky scores or the overall self-evaluation of the patients [p>0.05]
Conclusion: Thalidomide can effectively control the delayed vomiting of cancer patients receiving chemotherapy and the adverse reactions of the agent can be tolerated
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Objective To enrich breast cancer stem cells of breast cancer cell lines MCF-7 and MDA-MB-231 through culturing mammospheres,and to detect the expression differences of gene of breast cancer stem cells makers.Methods The MCF-7 and MDA-MB-231 cell lines were cultured by serum and serum-free medium.The proportion of CD44+/CD24-and CD133+ cancer stem cells was measured in cells derived from mammosphere cells or monolayer culture cells by flow cytometry,and the expression of CD44,CD24,CD133,ALDH3A1,ABCG2 and CXCR4 mRNA were detected by RT-PCR.Results Flow cytometry analysis indicated that the CD44+/CD24-low proportion of the MCF-7 mammosphere cells was higher than its adherent culture cells (P ＜ 0.05),and CD133+ proportion had no difference between them (P ＞ 0.05).However,the CD44+/CD24-/low proportion of the MDA-MB-231 mammosphere cells was lower than its adherent culture cells (P ＜ 0.05),while CD133+ proportion was significantly higher than its adherent cultured cells (P ＜ 0.05).RT-PCR analysis suggested that the expression of CD44 and ABCG2 increased obviously in MCF-7 microspheres (P＜ 0.05),and the expression of CD24,CD133,ALDH3A1 and CXCR4 had no significant difference between the mammosphere cells and adherent culture cells (P ＞ 0.05).The CD24,CD133,ABCG2,ALDH3A1 and CXCR4 increased obviously in MDA-MB-231 microspheres.On the contrary,the CD44 decreased (P ＜ 0.05).The expression of CD44 and CD24,CD133 and ALDH3A1 had significant differences in the microspheres of MCF-7 and MDA-MB-231 cells (P ＜ 0.05).Conclusion The related cancer stem cells gene expression is different in the microspheres of human breast cancer cell line,which indicates that the different subtypes of breast cancer may be derived from different origins.
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Objective:To investigate the effect on proliferation and apoptosis of T-cell leukemia cells by silencing NRP-1 ( Jurkat cells).Methods:The lentivirus plasmid which expresses NRP1 gene specific shRNA was constructed in our preliminary ex-perimental.We transfected the lentivirus plasmid to human T-cell Lymphoma cells.The proliferation of Jurkat cells different groups and effect on cell proliferation after chemotherapy drug EPI-treated were found by CCK-8 kit.The proliferation level and apoptosis rate of the cells were detected by flow cytometry and Annexin-V-FITC/PI method.Results:The proliferation level of NRP-1 /shRNA interference group was decreased significantly in 48 h,72 h,96 h,which was compared with the control groups.The apoptosis rate of the NRP-1/shRNA interference group was increased compared with control groups.The chemotherapy drug sensitivity of epirubicin ( EPI ) test results showed that EPI concentration was 0.025,0.05,0.1,0.2,0.4 μg/ml,the NRP-1/shRNA interference group of cell growth inhibition rate was increased,the corresponding control group difference had statistical significance(P<0.05).We choose the drug con-centration of the EPI IC50 for next experiments.NRP-1/shRNA interference group cell apoptosis rate increased significantly after induction,compared with the control groups difference was statistically significant ( P<0.05 ).Compared with control group, the expression level of Bcl-2 protein was decreased and the expression level of bax protein was increased significantly after EPI induction.The percentage of cells at G0/G1 phase increased significantly,while those at S phase decreased significantly.Conclusion:Plasmid shRNA-NRP1 inhibited the expression of NRP1 in Jurkat cells and decreased the proliferation level of Jurkat cells and promote their apoptosis and enhance their drug sensitivity;the molecular mechanism may relate to down-regulation of Bcl-2 and up-regulation of Bax.and arrested the cell cycle at G0/G1 phase.
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Objective:To construct GST-tagged human NRP-1 fusion protein expression vector and induce its expression in Escherichia coli ( E.coli) ,then carry on inclusion body refolding and purification so as to obtain GST-NRP-1 fusion protein.Methods:NRP-1 gene was amplified by RT-PCR and inserted into pCR-blunt vector.Then the reconstructed plasmid was inserted into prokaryotic expression vector pGEX-4T-1.The constructed pGEX-4T-1-NRP-1 expression vector was transformed into BL21 cells and induced by i-sopropyl-β-D-thiogalactoside ( IPTG).Bacterial bodies were disrupted by sonication.Then the soluble fraction of fusion proteins were verified by Western blot and purified by Glutathione Sepharose 4B after inclusion body refolding.Results: The NRP-1 gene fragment was amplified by RT-PCR and inserted into pCR-blunt vector.Fusion protein expression vector pGEX-4T-1-NRP-1 was constructed suc-cessfully.After transformation, GST-NRP-1 expression vector was detected in BL21 cells and obtained purifying protein after refolding.Conclusion:The plasmid GST-NRP-1 was constructed successfully and laid basis for subsequent studies.
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Objective To investigate the effect of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) combined with recombinant bovine basic fibroblast growth factor (rb-bFGF) in the treatment of oral ulcer caused by chemotherapy.Methods 108 patients of oral ulcer caused by chemotherapy were randomly divided into two groups.54 cases in the control group were treated with cydiodine buccal tablets at first,then received the aerosol treatment which was prepared by mixing gentamicin,dexamethasone,2 ％ lidocaine and physiologic saline,three times per day.54 cases in the treatment group firstly received the gargle which was prepared by mixing rhGM-CSF,dexamethasone and physiologic saline,then were treated with rb-bFGF by spraying on the oral ulcer surface,three times per day.Results The effective rate of the treatment group was 96.30 ％ (52/54),which was significantly higher than that of the control group [64.81％ (35/54)],there was a significant difference between the two groups (x2 =17.08,P ＜ 0.05).Conclusion The effect of rhGM-CSF combined with rb-bFGF in the treatment of oral ulcer caused by chemotherapy is very significant.
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Objective To explore the therapeutic effects and adverse reactions of thalidomide in the treatment of peri-chemotherapy nausea and vomiting of cancer patients.Methods Total of 70 patients were randomly divided into two groups:the treatment group (38 cases) and the control group (32 cases).The treatment group was treated with thalidomide (oral administration at a dose of 100 mg per night,then dose can be added by 50 mg until the top dose of 200 mg per day).The original will be maintained if they cannot be tolerate of extensive dose.The treatment group was also injected 2mg tropisetron in 30 minutes before chemotherapy.The control group was only injected same dose tropisetron.All cases were examined antiemetic effects and evaluated adverse reactions.Results Nausea and vomiting control rates were 89.5 ％ (34/38) and 68.8 ％ (22/32) respectively in the treatment group and control group respectively with significant difference.The adverse reactions were similar between the two groups.Conclusion Thalidomide joint tropisetron can effectively control the peri-chemotherapy nausea and vomiting,and the adverse reactions can be acceptable.It could improve further indications of the drug.
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Objective To construct the small hairpinRNA recombinant plasmids targeting mdr-1 gene which expresses highly in ovarian cancer resistance strain SKOV3/TAXOL to silence endogenetic mdr-1 gene expression and investigate the role of mdr-1 gene in the development of resistant ovarian cancer. Methods The pPGPU6/GFP/Neo-mdr-1 were constructed by gene clone technology. The influence on proliferation and apoptosis were investigated by CCK-8 in SKOV3/TAXOL after transfected. pPGPU6/GFP/Neo-mdr-1. Results The expression against mdr-1 proteins were inhibited by pPGPU6/GFP/Neo-mdr-1. The cell proliferation were inhibited after transfected pPGPU6/GFP/Neo-mdr-1 by CCK-8. The apoptosis were observed in DAB experiments and the apoptosis rate incised. Conclusion mdr-1 plays an important role in proliferation of resistant ovarian cancer and the short hairpinRNA of mdr-1 can efficiently suppress mdr-1expression and enhance the apoptosis in SKOV3/TAXOL, which provides a novel method for chemotherapy resistant tumors.
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Objective To construct the short hairpin RNA recombinant plasmids targeting mdr1 gene which expresses highly in ovarian cancer resistance strain SKOV3/TAXOL to silence endogenefic mdr1 gene expression and investigate the role of mdr1 gene in the development of resistant ovarian cancer. Methods The pGPU6/GFP/Neo-mdr1 were constructed by gene clone technology. The influence on proliferation and apoptosis were investigated by CCK-8 in SKOV3/TAXOL after transfected pGPU6/GFP/Neo-mdr1. Results The expression against mdr1 proteins were inhibited by pGPU6/GFP/Neo-mdr1. The cell proliferation were inhibited after transfected pGPU6/GFP/Neo-mdr1 by CCK-8. The apoptosis were observed in DAB experiments and the apoptosis rate increased. Conclusion mdr1 plays an important role in proliferation of resistant ovarian cancer and the short hairpin RNA of mdr1 can efficiently suppress mdr1 expression and enhance the apoptosis in SKOV3/ATAXOL.