Correlation Analysis of ADPRT rs1136410 Polymorphism with the Occurrence of Non-small Cell Lung Cancer in Han Nationa- lity from Northern Jiangsu / 中国药房

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.

Experimental study of PLCεon migration and invasion of human osteosarcoma cancer cells / 中国药理学通报

Aim To investigate the effects of PLCε on the invasion and migration of human osteosarcoma cancer cells U2OS. Methods RNA interference ( RNAi) was used to inhibit PLCεexpression, and the proliferation of cancer cells was measured by CCK-8 assay. The migration of the cells was measured by scratch wound healing assay and migration chamber as-say. Gelatin zymography was performed to measure the MMP2 activities in U2OS cells. Results PLCε ex-pression was suppressed by siRNA. CCK-8 assay showed that PLCε had no effect on the proliferation of cancer cells. PLCε knockdown inhibited cell invasion and activities of MMP2 . Conclusion PLCε knock-down can inhibit the migration and invasion of human osteosarcoma cancer cells U2OS.

Role of glutamate receptor-6 in kainate-induced epilepsy in rats / 西安交通大学学报(医学版)

Objective To study the molecular mechanism of glutamate receptor-6 (GluR6) in the pathogenesis of epilepsy. Methods Seizure model of SD rats was induced by intraperitoneal injection of kainate (KA). Immunoprecipitation and immunoblotting were performed to examine the interactions of GluR6 and MLK3 with PSD95 at various time points after KA injection. The effect of Tat-GluR6-9c on the MLK3 phospharylation induced by kainate was observed with immunoblotting and immunohistochemistry. Results The assembly of GluR6 and MLK3 with PSD95 was induced after KA hippocampal CA3 region, and bagan to decrease one day later. Pretreatment after KA injection in CA3 region (P<0.05). Conclusion KA induces the assembly of the GluR6-PSD95-MLK3 signaling module and subsequently activates MLK3, which ultimately results in brain injury.

Oncolytic adenoviruses harboring IL-24 gene induce the apoptosis of melanoma cells / 中华皮肤科杂志

Objective To study the effects of oncolytic adenoviruses ZD55 harboring IL-24 gene (ZD55-IL-24) on the apoptosis of human melanoma cell line A375. Methods The oncolytie adenoviruses ZD55-IL-24 were verified by PCR. Then, the viruses were propagated, purified, and titrated by HEK293 cell plaque assay. A375 cells were cultured, divided into three groups transfected with ZD55-1L-24, ZD55 fused with enhanced green fluorescent protein (ZD55-EGFP), and replication-deficient adenovirus ZD55 carrying IL-24 gene (AD-IL-24), respectively. The multiplicity of infection was 0.1, 1, 10 and 100, respectively.Subsequently, the eytotoxity of these viruses and proliferation of A375 cells were determined by crystal violet staining and methyl thiazolyl tetrazolium (MTT) assay, respectively. The expressions of EIA and IL-24 protein were detected by Western blot in A375 cells. Results PCR verified that the adenoviruses ZD55-IL-24 contained IL-24 gene without wild adenovirns contamination. Crystal violet staining revealed that ZD55-IL-24 had an obvious eytotoxic effect on A375 cells, and MTT assay indicated that ZD55-IL-24 inhibited the proliferation of A375 cells in a time-and concentration-dependent manner. As shown by Western blot analysis, ZD55-1L-24 expressed IL-24 and E1A protein in A375 cells with a high efficiency. Conclusions The oncolytic adenoviruses ZD55-IL-24 can efficiently express IL-24 gene, inhibit the proliferation of, and induce the apoptosis in A375 cells.

Antitumor effects of oncolytic adenovirus expressing human IL-18 gene on malignant melanoma implanted in nude mice / 中华皮肤科杂志

Objective To evaluate the antitumor effect of oncolytic adenovirus expressing human IL-18 gene on malignant melanoma implanted in nude mice.Methods BALB/c nude mice were subcutaneously inoculated with A375 cells to establish a model of malignant melanoma.When the volume of implanted tumor reached 100-150 mm3,murine models were randomly divided into three groups to receive a 3-day intratumoral injection of IL-18 gene-expressing human oncolytic adenovirus named ZD55-IL-18,IL-18 gene-expressing adenovirus named Ad-IL-18,phosphate buffer saline(PBS),respectively.The tumor size was measured at an interval of 4 days for 9 weeks.Hematoxylin and eosin (HE)staining was performed to observe the morphological changes of tumor cells.The protein expression of IL-18 and E1A.microvessel density in tumor tissue,and apoptosis of tumor xenografts were detected by immuno fluorescence assay,immunohistochemistry and in situ end labeling technique (TUNEL).respectively.Results The treatment with ZD55-IL-18 significantly inhibited the growth of tumor.Forty-four days after the treatment,the mean tumor volume was 1039.378±29.67 mm3 in ZD55-IL-18-treated mice.significantly smaller than that in Ad-IL-18 treated mice(2900.46±62.65 mm3)and PBS-treated mice(3980.24±63.78 mm3).HE staining showed that the nuclei of tumor cells were heavily stained with few nucleoli in ZD55-IL-18-treated mice.Increased positivity rate of IL-18 was noticed in ZD55-IL-18-treated mice vs.AD55-IL-18-treated mice(83.4%±3.2%vs 24.4%±2.1%.P＜0.01).Moreover,immunofluorescence assay revealed the presence of E1A protein in tumor tissue.A decrease was found in the microvessel density in ZD55-IL-18-treated mice compared with the PBS-treated mice(P＜0.01).The apoptosis rate in tumor cells from high to low was 86.28%±3.25%in ZD55-IL-18-treated mice,43.67%±3.46%in Ad-IL-18-treated mice,and 10.73%±2.48%in PBS-treated mice;there was a significant difference between the three groups(all P＜0.05).Conclusion The oncolytic adenovirus expressing human IL-18 gene,ZD55-IL-18,has a significant inhibitory effect on the growth and metastasis of malignant melanoma implanted in nude mice.