目的 探讨干扰LINC00707对食管癌细胞生物行为的影响及分子机制.方法 选取51例食管癌患者癌组织及癌旁正常组织,用实时荧光定量-聚合酶链反应(RT-qPCR)检测LINC00707和miR-382-5p的表达水平;将食管癌细胞EC9706随机分为对照(con)组、si-LINC00707组、si-NC组、miR-382-5p组、miR-NC组、anti-miR-382-5p组、anti-miR-NC组、si-LINC00707+anti-miR-382-5p组;用四甲基偶氮唑盐(MTT)比色法检测细胞活性;克隆形成实验检测细胞克隆形成数;流式细胞术检测细胞凋亡率;划痕实验检测细胞迁移距离;Western印迹法检测蛋白表达;双荧光素酶报告实验检测LINC00707和miR-382-5p的靶向关系.结果 与癌旁正常组织相比,食管癌组织中LINC00707表达水平升高,miR-382-5p表达水平降低,差异有统计学意义(P<0.05);且LINC00707和miR-382-5p表达水平呈负相关(P<0.05).干扰LINC00707或过表达miR-382-5p后,EC9706细胞活性降低,克隆形成数减少,细胞凋亡率升高,细胞迁移距离缩短,Bax表达水平升高,Bcl-2表达水平降低,差异有统计学意义(P<0.05).抑制miR-382-5p表达与过表达miR-382-5p对EC9706细胞的作用相反.且抑制miR-382-5p表达逆转了干扰LINC00707对EC9706细胞的作用.LINC00707靶向调控miR-382-5p.结论 干扰LINC00707可能通过上调miR-382-5p抑制食管癌细胞的恶性生物学行为.

Objective:To investigate the relationship between the serum alkaline phosphatase (ALP), prostate specific antigen (PSA) and bone metastasis in initially diagnosed prostate cancer (PCa) patients in different ISUP(International Society of Urological Pathology)groups.Methods:The 368 initial diagnosed prostate cancer patients recruited from January 2013 to December 2018 were retrospectively analyzed, including 247 cases in the Third Affiliated Hospital of Sun Yat-sen University, 111 cases in the Yuebei People's Hospital Affiliated to Medical College of Shantou University and 10 cases in Shenzhen Hospital of Southern Medical University. According to whether there was bone metastasis at the initial diagnosis, it was divided into 230 cases in the bone metastasis group and 138 cases in the non bone metastasis group. There was no significant difference between the two groups in age [(71.9±9.4) years and (71.2±8.7) years], body mass index (BMI) [(23.1±3.7) kg/m 2 and (23.7±2.6) kg/m 2]. There were significant differences in PSA [(307.3±847.0) ng/ml and (84.5±257.3) ng/ml] and ALP [(174.5±270.8) U/L and (71.0±23.2) U/L] between the two groups. In different PSA subgroups, there were 45 cases in PSA <10 ng/ml, 35 cases in PSA 10-20 ng/ml and 288 cases in PSA >20 ng/ml. The differences of ALP and PSA between bone metastasis group and non-bone metastasis group based on different ISUP stratification were analyzed, ROC curves were used to predict their risks of bone metastasis. Results:There were 3(1.3%), 22(9.6%), 34(14.8%), 85(37.0%) and 86 (37.4%) prostate cancer patients with bone metastasis from ISUP group 1 to 5, and 14(10.1%), 19(13.8%), 29(21.0%), 32(23.2%) and 44(31.9%) without bone metastasis, respectively. There was significant difference in the serum ALP levels between the bone metastasis group and the boneless metastasis group in the ISUP group 4(157.6±207.7 vs. 66.5±17.0) and 5(189.4±257.5 vs. 69.2±18.4)( P<0.001) and PSA levels had difference in the ISUP group 3(240.3±313.0 vs. 42.4±42.1), 4(152.3±184.5 vs. 44.7±33.3) and 5(435.2±1006.3 vs. 60.8±84.8)( P<0.001). There was statistically significant between the bone metastasis group and the without(336.1±882.2 vs. 139.3±328.1) when PSA>20 ng/ml( P=0.006). ROC curve analysis: the cut-off values of ALP were 115.5, 109.0, 75.5 and 86.0 U/L from ISUP group 2 to 5 respectively, the sensitivity was 23.8%, 56.5%, 66.4% and 50.6% respectively, the specificity was 99.7%, 93.4%, 78.3% and 89.2% respectively, and the accuracy were 59.4%, 73.1%, 69.7% and 63.3%, respectively. The cut-off values of PSA were 39.5, 93.1, 54.2 and 28.9 ng/ml from ISUP group 2 to 5 respectively, the sensitivity was 64.4%, 68.4%, 87.4% and 88.3% respectively, and the specificity was 79.5%, 90.6%, 63.7% and 61.6% respectively, and the accuracy were 71.6%, 78.1%, 80.1% and 79.2%, respectively. Conclusion:ALP increased significantly in ISUP group ≥4 and PSA in ISUP group ≥3, which related to bone metastasis in patients with initial diagnosed prostate cancer.

Objective To explore the effects of hsa-miR-96 to A549. Method A549 were transfected with miR-96 inhibitor/mimics and shRNA-mTOR. The proliferation were detected by MTT,the expression of p21, cyclin D1,p-4E-BP,S6K were detected via RT-PCR and WB. The luciferase assay were used to anaylse the target of miR-96. Result With or without metformin treated,up-regulated the expression of miR-96 could inhibit A549 proliferation,increase p21 expression and decrease cyclin D1 expression instead. The results may related with G1 arrest which induced by miR-96 up-regulation. The ratio of firefly fluorescence value/Renilla fluorescence value was lower than that in wild type NC or mutant group. Conclusion miR-96 target with mTOR to inhibit the growth of A549.

目的 观察肺癌组织 Notch1、Bmi-1蛋白表达变化,探讨检测肺癌组织 Notch1、Bmi-1蛋白表达的临床意义.方法 选择67例肺癌组织标本为观察组,30例行肺支气管镜检查者的正常肺组织标本为对照组.用免疫组化链霉菌抗生物素蛋白-过氧化物酶连结法(S-P法)检测两组Notch1、Bmi-1蛋白表达.分析肺癌组织 Notch1、Bmi-1蛋白表达与肺癌临床病理参数的关系.结果 观察组、对照组Notch1阳性表达率分别为62.69%(42/67)、13.33%(4/30),Bmi-1阳性表达率分别为49.25%(33/67)、10.00%(3/30).观察组Notch1、Bmi-1蛋白阳性表达率均高于对照组,P均<0.05.肺癌组织Notch1、Bmi-1表达与淋巴结转移、分化程度、临床分期有关(P均<0.05),与年龄、性别、病理类型无关(P均>0.05).结论 肺癌组织Notch1、Bmi-1表达升高.检测肺组织Notch1、Bmi-1表达有助于肺癌的诊断及病情评估.

[目的]探讨食管鳞癌患者围手术期外周血中的循环肿瘤细胞(CTC)检出水平的意义及预后价值.[方法]检测48例食管鳞癌患者术前1 d、术后7 d外周静脉血CTC,分析术前CTC检出阳性水平与临床病理生理特征的关系,分析术前、术后CTC检出阳性水平与术后无进展生存时间(PFS)的关系.[结果]术前CTC不同检出水平与细胞分化程度、淋巴结转移状态及p- TNM分期关系密切.肿瘤分化程度越低(χ2=9.636,P=0.008)、淋巴结转移越多(χ2=4.218,P=0.040)、p-TNM分期越晚(χ2=8.392,P=0.004),CTC检出阳性率越高.术前、术后血CTC检出阳性组的术后PFS(均为20个月)较术前、术后检出阴性组低(均为13个月),且差异均有显著性(均P<0.01).[结论]食管鳞癌患者外周血CTC的检出水平可以反应肿瘤的进展程度及作为预后判断的指标,CTC水平越高可能提示预后不良.

AIM:To express N terminal fragment of human Toll-like receptor-9 (hTLR-9) in E.coli and prepare its antiserum.METHODS:The thioredoxin fusion expression plasmid of terminal fragment of hTLR-9 gene (707-1 167 bp) was constructed and then transformed into E.coli BL21 (DE3). The fusion protein was expressed in E.coli BL21 (DE3) under induction of 0.5 mmol/L IPTG. Inclusion body dissolved in urea was used as immunogen to prepare mouse anti-hTLR-9 antibody.RESULTS:The fusion protein was expressed in the form of inclusion body with molecular mass of about 31kD. The purity of fusion protein reached to 90%; The result of immunohistochemical staining showed that the antisera against recombinant fusion protein could react specifically to HEK293 cells transfected with hTLR-9.CONCLUSION:The fusion protein of hTLR-9 N-terminal fragment was expressed in E.coli BL21 (DE3). The prepared anti-hTLR-9 antibody can react specifically to HEK293 cells transfected with full length hTLR-9.