Expression and clinical significance of dendritic cell and transforming growth factor-beta 1 in cervical cancer

Objective:To explore the density and mature status of Dendritic cell(DC) in cervical cancer and correlation with the expression of transforming growth factor-beta 1(TGF-β1).Methods:Streptavidin-peroxidase(SP) immunohistochemistry methods were used to detect S-100 DC and the expression of TGF-β1 in 20 normal cervical tissues and 53 cervical cancer tissues without any sort of chemotherapy or radiation therapy prior to resection.Medical records were reviewed,clinicopathological variables were retrieved and used for analysis.Results:Two types of DC were observed under the microscope.The expression of DC in cervical cancer was significantly higher than that in normal tissues(23.34 cells/mm^2 vs 29.91 cells/mm^2,P<0.05),and significantly higher in early stage than that in advanced stage(P<0.05).The expression of TGF-β1 was significantly higher in cervical cancer than that in normal tissues (P<0.025).However,there was no correaction between TGF-β1 and lymph nodes metastasis.The index of DC in cervical cancer was negatively correlated to the expression of TGF-β1 in tumor cells (r=-0.8875,P=0.0001).Conclusion:Maturation of DC in cervical cancer is inhibited.The decreased number of DC and the higher expression of TGF-β1 are due to the failure of the immunity,these may play an important role in the development of the cervical cancer.

Expression of vascular endothelial growth factor and its prognostic significance in gastric carcinoma

AIMS To investigate the clinical significance of vas- cular endothelial growth factor (VEGF) expression in gastric carcinoma. METHODS The expression of VEGF in 128 gastric carcinomas was investigated by immunohistochemical staining with a polyclonal antibody against VEGF. Cor- relations between the expression of VEGF and various clinicopathologic factors and prognosis were studied. RESULTS The VEGF-rich expression rate was 64.1% in gastric carcinoma. VEGF-rich expression rate of patients with stage Ⅲ and stage Ⅳ disease was greater than that of patients with stage f disease (P <0.05). Significant differences of expression rate ex- isted with respect to growth pattern,serosal invasion and lymph node metastasis. The VEGF-rich rate was much higher in tumors with expanding growth pattern (71.8%) or serosal invasion (73.5%) than in those with infiltrative growth pattern (52.0%) or non-serosal invasion (53.3%) (P<0.025,respectively),and it was also significantly higher in patients with lymph node metastases (75.0%) than in those without such metastases (50.0%) (P<0.05). In addition,postop- erative survey of 86 patients who had been followed up for at least 5 years demonstrated that the 5-year sur- vival rate of patients with VEGF-rich tumors was signifi- cantly lower than that of patients with VEGF-poor tu- mors (P<0.05). CONCLUSIONS The expression of VEGF may be as- sociated with the invasion and metastasis and may also be a useful prognostic indicator of gastric carcinoma.

Expression of vascular endothelial growth factor and its role in oncogenesis of human gastric carcinoma

AIM: To establish the role of vascular endothelial growth factor (VEGF) in the oncogenesis of human gastric carcinoma more directly. METHODS: The expression of VEGF and its receptor kinase-domain insert containing receptor (KDR) in human gastric cancer tissue were observed by immunohistochemical staining. VEGF levels were manipulated in human gastric cancer cell using eukaryotic expression constructs designed to express the complete VEGF(165) complimentary DNA in either the sense or antisense orientation. The biological changes of the cells were observed in which VEGF was up-regulated or down-regulated. RESULTS: VEGF-positive rate was 50%, and VEGF was mainly localized in the cytoplasm and membrane of the tumor cells, while KDR was mainly located in the membrane of vascular endothelial cells in gastric cancer tissues and peri-cancerous tissue. In 2 cases of 50 specimens, the gastric cancer cells expressed KDR, localized in both the cytoplasm and membrane. Introduction of VEGF(165) antisense into human gastric cancer cells (SGC-7901, immunofluorescence intensity, 31.6%)) resulted in a significant reduction in VEGF-specific messenger RNA and total and cell surface VEGF protein (immunofluorescence intensity, 8.9%) (P【0.05). Conversely, stable integration of VEGF(165) in the sense orientation resulted in an increase in cellular and cell surface VEGF (immunofluorescence intensity, 75.4%) (P【0.05). Lowered VEGF levels were associated with a marked decrease in the growth of nude mouse xenografted tumor (at 33 days postimplantation, tumor volume: 345.40 +/- 136.31 mm3)(P【0.05 vs control SGC-7901 group: 1534.40 +/- 362.88 mm3), whereas up-regulation of VEGF resulted in increased xenografted tumor size (at 33 days postimplantation, tumor volume: 2350.50 +/- 637.70 mm3) (P【0.05 vs control SGC-7901 group). CONCLUSION: This study provides direct evidence that VEGF plays an important role in the oncogenesis of human gastric cancer.

Correlation of integrin β3 mRNA and vascular endothelial growth factor protein expression profiles with the clinicopathological features and prognosis of gastric carcinoma

AIM： To investigate integrin β3 mRNA and vascular endothelial growth factor （VEGF） protein expression in gastric carcinoma, and its correlation with microvascular density, growth-pattern, invasion, metastasis and prognosis. METHODS： In situ hybridization（ISH） of integrin β3 mRNA and immunohistochemistry of VEGF and CD34 protein were performed on samples from 118 patients with gastric cancer. RESULTS： The positive rate of integrin β3 mRNA in non-tumor gastric mucosa （20%） was significantly lower than that of the gastric cancer tissue （52.5%, X^2 = 10.20, P 〈 0.01）. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the positive expression rates of integrin β3 mRNA were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the positive expression rates of VEGF protein were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the mean MVD were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. It was found that the positive expression rate of integrin β3 mRNA was positively related to that of VEGF protein （P 〈 0.01） and MVD （P 〈 0.05）, meanwhile the positive expression rate of VEGF protein was positively related to MVD （P 〈 0.05）. The mean survival period in patients with positive expression of integrin β3 mRNA and VEGF, and MVD ≥54.9/mm^2 was significantly shorter than that in patients with negative expression of integrin β3 mRNA （P 〈 0.05） and VEGF （P 〈 0.01）, and MVD 〈 54.9/mm^2 （P 〈 0.01）. Five-year survival rate in patients with positive expression of integrin β3 mRNA and VEGF, and MVD ≥54.9/mm^2 was significantly lower than those with negative expression of integrin β3 mRNA （P 〈 0.05）, VEGF （P 〈 0.05）, and MVD 〈 54.9/mm^2 （P 〈 0.01）. CONCLUSION： Integrin β3 and VEGF expression can synergistically enhance tumor angiogenesis, and may play a crucial role in invasion and metastasis of gastric carcinoma. Therefore, they may be prognostic biomarkers and novel molecular therapeutic targets.

Effects of epidermal growth factor on the growth of human gastric cancer cell and the implanted tumor of nude mice

AIM: Epidermal growth factor (EGF) plays an important role in the regulation of gastrointestinal tissue growth and development, and it can stimulate epithelial proliferation, cell differentiation and growth. It has been established that the EGF can promote gastric cytoprotection and ulcer healing. But the potential ability of EGF to regulate the gastric cancer growth is unknown. This study is to investigate the influence of EGF on human gastric cancer cell and the implanted tumor growth of nude mice. METHODS: The cell growth rates of human gastric adenocarcinoma cell lines MKN-28, MKN-45, SGC-7901 and normal human gastric epithelial cells 3T3 were assessed when incubated with recombinant human EGF (rhEGF, 0.05, 0.1, 0.5, 1.0, 10, 50, 100 mg.L(-1)) using MTT method. The cells of MKN-28, MKN-45, SGC-7901 (gastric cancer tissue 1.5mm(3)) were implanted in the BALB/cA nude mice for 10 days.The EGF was given intraperitoneally (15, 30, 60 microg.kg(-1)) for 3 weeks. The body weights of the tumor-bearing animals and their tumor mass were measured afterwards to assess the mitogenic effect of rhEGF in the nude mice. RESULTS: Within the concentration range of 0.05-100mg.L(-1), rhEGF could increase the cell growth of normal 3T3 cells (cell growth rate 100% vs 102.8%, P【0.05), but partially restrain the gastric cancer cell growth. The latter effect was related to cell differentiation. In 15-60 microg/kg rhEGF groups, the mean implanted tumor mass of MKN-28 cell were 1.75 g, 1.91 g, 2.08 g/NS group 1.97 g (P】0.05), the mean tumor mass of SGC-7901 cell were 1.53 g, 1.07 g, 1.20 g/NS group 1.07 g (P】0.05), and for MKN-45 cell, the tumor mass were respectively 1.92 g, 1.29 g, 1.77 /NS group 1.82 g (P】0.05). So rhEGF had no obvious effect on implanted MKN-28, SGC-7901 and MKN-45 tumor growth. CONCLUSION: EGF has no stimulating effect on the human gastric cancer cell growth neither in vitro nor in vivo.

Correlative studies on uPA mRNA and uPAR mRNA expression with vascular endothelial growth factor, microvessel density, progression and survival time of patients with gastric cancer

AIM： To investigate the correlations between the expression of urokinase-type plasminogen activator （uPA） mRNA, uPA receptor （uPAR） mRNA and vascular endothelial growth factor （VEGF） protein and clinicopathologic features, microvessel density （MVD） and survival time. METHODS： In situ hybridization and immuno-histochemistry techniques were used to study the expressions of uPA mRNA, uPAR mRNA, VEGF and CD34 protein in 105 gastric carcinoma specimens. RESULTS： Expressions of uPA mRNA, uPAR mRNA and VEGF protein were observed in 61 （58.1%） cases, 70 （66.7%） cases and 67 （63.8%） cases, respectively. The uPA mRNA and uPAR mRNA positive expression rates in infiltrating-type cases （73.7%, 75.4%）, stage Ⅲ- Ⅳ （72.1%, 75.4%）, vessel invasion （63.2%, 69.9%）, lymphatic metastasis （67.1%, 74.4%） and distant metastasis （88.1%, 85.7%） were significantly higher than those of the expanding-type （χ^2 = 15.57, P = 0.001; χ^2 = 6.91, P = 0.046）, stage Ⅰ-Ⅱ （χ^2 = 19.22, P = 0.001; χ^2 = 16.75, P = 0.001）, non-vessel invasion （χ^2 = 11.92, P = 0.006; χ^2 = 14.15, P = 0.002）, non- lymphatic metastasis （χ^2 = 28.41, P = 0.001; χ^2 = 22.5, P = 0.005） and non-distant metastasis （χ^2 = 12.32, P = 0.004; χ^2 = 17.42, P = 0.002; χ^2 = 11.25, P = 0.012; χ^2 = 18.12, P = 0.002）.The VEGF positive expression rates in infiltrating-type cases （75.4%）, stage Ⅲ-Ⅳ （88.5%）, vessel invasion （82.9%）, lymphatic metastasis （84.3%） and distant metastasis （95.2%） were significantly higher than those of the expanding-type （χ^2 = 9.61, P = 0.021）,stage Ⅰ-Ⅱ （χ^2 = 16.66, P = 0.001）, non-vessel invasion （χ^2 = 29.38, P = 0.001）, non-lymphatic metastasis （χ^2 = 18.68, P = 0.005）, and non-distant metastasis （χ^2 = 22.72, P = 0.007; χ^2 = 21.62, P = 0.004）. The mean MVD in the specimens positive for the uPA mRNA, uPAR mRNA and VEGF protein was markedly higher than those with negative expression groups. Moreover, a positive relation between MVD and uPA mRNA （rs = 0.199, P = 0.042）, uPAR mRNA （rs = 0.278, P = 0.035）, and VEGF （rs = 0.398, P = 0.048） expressions was observed. The mean survival time in cases with positive uPA mRNA, uPAR mRNA and VEGF protein expression or MVD value ≥54.9 was significantly shorter than those in cases with negative expression or MVD value 〈 54.9. CONCLUSION： uPA and uPAR expressions are correlated with enhanced VEGF-induced tumor angiogenesis and may play a role in invasion and nodal metastasis of gastric carcinoma, thereby serving as prognostic markers of gastric cancer.

C-reactive protein,procalcitonin,interleukin-6,vascular endothelial growth factor and oxidative metabolites in diagnosis of infection and staging in patients with gastric cancer

AIM:The current study was to determine the serum/pLasma levels of VEGF,IL-6,malondialdehyde (MDA),nitric oxide (NO),PCT and CRP in gastric carcinoma and correlation with the stages of the disease and accompanying infection. METHODS:We examined the levels of serum VEGF,IL-6, PCT,CRP and plasma MDA,NO in 42 preoperative gastric cancer patients and 23 healthy subjects.There were infection anamneses that had no definite origin in 19 cancer patients. RESULTS:The VEGF levels (mean±SD; pg/mL) were 478.05±178.29 and 473.85±131.24 in gastric cancer patients with and without infection,respectively,and these values were not significantly different (P>0.05).The levels of VEGF, CRP,PCT,It-6,MDA and NO in cancer patients were significantly higher than those in healthy controls and the levels of CRP,PCT,It-6,MDA and NO were statistically increased in infection group when compared with non- infection group (P<0.001). CONCLUSION:Although serum VEGF concentrations were increased in gastric cancer,this increase might not be related to infection.CRP,PCT,IL-6,MDA and NO have obvious drawbacks in the diagnosis of infections in cancer patients. These markers may not help to identify infections in the primary evaluation of cancer patients and hence to avoid unnecessary antibiotic treatments as well as hospitalization. According to the results of this study,IL-6,MDA,NO and especially VEGF can be used as useful parameters to diagnose and grade gastric cancer.

Expression of c-erbB-2 oncogene protein, epidermal growth factor receptor, and TGF-β1 in human pancreatic ductal adenocarcinoma

Objective: To detect the relations of c-erbB-2 onco-gene protein, epidermal growth factor receptor (EG-FR) and transforming growth factor-β1 (TGF-β1)to the progression or metastasis of pancreatic carci-noma.Methods: Using streptavidinbiotin complex (SABC)method, c-erbB-2 oncongene protein, we examinedimmunohistochemically EGFR and TGF-β1 expres-sions in wax-tissue sections from 10 individuals withnormal pancreas (NP), 13 patients with chronic pan-creatitis (CP) and 36 patients with pancreatic ductaladenocarcinoma (PC).Results: The positive expression rates of c-cerbB-2oncogene protein, EGFR and TGF-β1 in the NP, CPand PC groups were 0, 0, 10%; 7.7%, 7.7%,7.7%; and 41.7%, 50.0%, 44.4%, respectively.The positive expression rates of the three specific pro-teins increased more significantly in the PC groupthan in the NP and CP groups (P【0.05). The indi-vidual expression of c-erbB-2, EGFR and TGF-β1was not related to the age and sex of the patients aswell as the site, size and histopathological grade oftumors (P】0.05), but to the clinical stage of tumors(P【0.01). The coexpression rate of the three pro-teins was 27.8 % (10/36). This coexpression in thePC group was correlated with the histopathologicalgrades and clinical stages of tumors (P【0.01).Conclusion: Detection of c-erbB-2 oncogene protein,EGFR, and TGF-β1 expressions in pancreatic tissueis helpful to judge the malignancy, progression, andmetastasis of PC.

Effects of Exogenous Growth Hormone on Growth Hormone-Insulin-Like Growth Factor Axis of Human Gastric Cancer Cell

Aim: To study effects of recombinant human growth hormone (rhGH) on growth hormone-insulin-like growth factor axis (GH-IGFs) of human gastric cancer cell in vivo in order to reveal part mechanism of growth effects of rhGH on gastric cancer. Methods: Nude mice were randomly divided into control group, cisplatin (DDP) group, rhGH group and DDP + rhGH group after human gastric cancer xenograft model of node mice was successfully founded and drugs were used for 6 days. We investigated volume of tumor, inhibitory rate of tumor and cell cycle by slide gauge and flow cytometry. In addition, We also respectively investigated insulin-like growth factor-I (IGF-I) and insulin-like growth factor binding protein-3 (IGFBP-3) of blood serum of nude mice, IGF-ImRNA, insulin-like growth factor-I receptor (IGF-IR) mRNA and IGFBP-3 mRNA of xenograft of nude mice by enzyme linked immunosorbent assay (ELISA) and semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) on the first day of completing use of drugs later. Results: Tumor grew obviously slowly and tumor inhibitory rate obviously rose in DDP group and DDP + rhGH group compared with control group and rhGH group (p p p < 0.05). Expressions of IGF-I mRNA and IGF-IR mRNA were not obviously different in all groups. But expression of IGFBP-3 mRNA obviously increased in rhGH group, DDP group and DDP + rhGH group compared with control group;meanwhile, expression of IGFBP-3 mRNA also obviously increased in DDP + rhGH group compared with control group, DDP group and rhGH group. Conclusion: Our results indicated rhGH in short-time use did not improve proliferation of human gastric cancer cells and its mechanism was possible that rhGH in short-time use raised simultaneously IGF-I and IGFBP-3 of blood serum and increased IGFBP-3 mRNA, but degraded ratio of IGF-I and IGFBP-3 of blood serum in human gastric cancer cells.

Vascular endothelial growth factor promotes angiogenesis in gastric carcinoma

Objective: To explore the role of vascular endothelial growth factor (VEGF) in the angiogenesis and development of human gastric carcinoma. Methods: The expressions of VEGF and its receptor KDR (ki-nase-domain insert containing receptor) in human gastric cancer tissue and SGC-7901 cells were detected with immunohistochemical staining. Microvessel density (MVD) was obtained after immunostaining for Factor-VIII. VEGF in SGC-7901 cell line was detected with Western blot. VEGF levels were manipulated in human gastric cancer cell by using eukaryotic expression vector containing the complete VEGF165 complimentary DNA in either the sense or antisense orientation. Finally the biological characteristics of the transfectants were identified. Results: VEGF-positive rate in TNM grade I and IV gastric carcinomas (19. 0%) were significantly higher than that in grade I and I (72. 4%) (P<0. 05). Increased MVD was found in VEGF-positive tumors (16. 4±6. 7). which is significantly larger than in VEGF-negative tumors (6. 5±2. 1) (P< 0. 05). Human gastric cancer cells (SGC-7901) produced 3 kinds of VEGF in molecule. In 2 cases of 50 specimens, a few gastric cancer cells expressed KDR in cytoplasm and cell membranes. SGC-7901 cells with anti-sense VEGF165 showed a significant reduction in cell surface VEGF protein with the immunofluorescence intensity from 8. 9% to 31. 6% (P<0. 05). However, those with stable integration of VEGF165 in the sense orientation resulted in an increase in cellular and cell surface VEGF with the immunofluorescence intensity from 75. 4% to 31. 6% (P<0. 05). The decrease of VEGF levels was associated with a marked decrease in the growth of nude mouse xenografted tumor (33 d post-implantation, 345. 4±136. 3 mm3 in size) (P<0. 05 vs control SGC-7901 group) , whereas VEGF overexpression resulted in an increase of xenografted tumor size (33 d post-implantation, 2 350. 5±637. 7 mm3 in size) (P<0. 05 vs control SGC-7901 group). Conclusion: VEGF plays an important role in the development of human gastric cancer, and might have an autocrine effect upon the gastric cancer cells. The inhibition of VEGF by antisense RNA expression might prevent solid tumor from growing and metastasizing.

miR-223-3p通过靶向TGFBR3促进LncRNA ADAMTS9-AS2表达上调抑制肺癌细胞的增殖和迁移作用

目的:探讨miR-223-3p通过靶向转化生长因子-βⅢ型受体(TGFBR3)促进长链非编码RNA(LncRNA)ADAMTS9-AS2表达上调抑制肺癌细胞增殖和迁移的作用及可能机制。方法:采用RT-PCR检测肺癌H1299细胞中miR-223-3p和TGFBR3 mRNA表达水平,Western blotting检测TGFBR3的蛋白表达水平,RT-qPCR检测LncRNA ADAMTS9-AS2的表达水平,CCK-8检测细胞增殖能力,Transwell细胞迁移实验和划痕实验检测细胞迁移能力。采用脂质体转染miR-223-3p模拟物(过表达组)、抑制剂(抑制组)、对照质粒(对照组)于H1299细胞中,比较各组转染前后miR-223-3p、TGFBR3、LncRNA ADAMTS9-AS2表达水平、细胞增殖能力、细胞迁移能力。结果:与转染前比较,过表达组转染后的H1299细胞中miR-223-3p、LncRNA ADAMTS9-AS2表达水平均升高(P<0.05),TGFBR3蛋白和mRNA表达水平均降低(P<0.01和P<0.05),细胞增殖和迁移能力下降(P<0.05);抑制组转染后的细胞中miR-223-3p和LncRNA ADAMTS9-AS2表达水平均降低(P<0.05),TGFBR3蛋白和mRNA表达水平均升高(P<0.01和P<0.05),细胞增殖和迁移能力均增加(P<0.05)。转染72 h后,TGFBR3蛋白表达水平及mRNA的表达水平细胞增殖与迁移能力:抑制组>观察组>过表达组(P<0.01)。3组miR-223-3p、LncRNA ADAMTS9-AS2 mRNA表达水平逐渐降低,抑制组<对照组<过表达组(P<0.01)。结论:miR-223-3p抑制肺癌H1299细胞的增殖和迁移,靶向下调TGFBR3和上调LncRNA ADAMTS9-AS2表达可能为其作用机制。

miR-25靶向BAMBI基因调控TGF-β信号通路对胃癌细胞增殖和转移的影响

目的探讨miR-25靶向骨形成蛋白和激活素的跨膜抑制剂(BAMBI)基因调控转化生长因子-β(TGF-β)信号通路对胃癌细胞增殖和转移的影响。方法选取2022年1月至2023年1月新疆医科大学第二附属医院收治的胃癌患者作为研究对象。收集胃癌患者胃癌组织为胃癌组,同时收集癌旁组织(远离病灶2 cm)作为癌旁组,每组30个样本。采用RT-PCR检测miR-25基因的表达,蛋白质印迹法检测及免疫组织化学检测BAMBI、TGF-β蛋白表达情况。胃癌SGC-7901细胞株构建体外模型,通过转染miR-25抑制慢病毒载体构建细胞模型。并依据处理方法的不同,分为对照孔、miR-25干扰慢病毒组和miR-25-NC组,评估细胞的迁移、侵袭、增殖能力及凋亡水平,并检测BAMBI、TGF-β蛋白的相对表达水平。结果与癌旁组相比,病灶组中miR-25基因表达量升高,BAMBI、TGF-β蛋白表达量降低,差异均有统计学意义(P<0.05)。与对照组、miR-25-NC组相比,miR-25干扰慢病毒组细胞迁移率、侵袭能力、细胞的增殖率降低,细胞的凋亡率升高,差异均有统计学意义(P<0.05)。对照组、miR-25干扰慢病毒组、miR-25-NC组中BAMBI表达量比较,差异无统计学意义(P>0.05);与对照组相比,miR-25干扰慢病毒组中TGF-β表达量升高,差异有统计学意义(P<0.05);对照组与miR-25-NC组TGF-β表达量比较,差异无统计学意义(P>0.05)。结论胃癌组织中miR-25/BAMBI/TGF-β信号通路被激活,且miR-25/BAMBI/TGF-β信号通路的激活可促进胃癌细胞的增殖、迁移、侵袭,并抑制凋亡凋亡,从而影响胃癌的预后发展。

联合多期增强CT纹理分析及血液学指标术前预测胃癌VEGFR2表达状态

目的探讨多期动态增强CT纹理分析参数联合血液学指标术前预测胃癌血管内皮生长因子受体2(vascu‐lar endothelial growth factor receptor2,VEGFR2)表达状态的价值。方法选取本院148例胃癌患者资料,获得术前血液学指标和三期增强CT纹理分析参数。基于组内相关系数和差异性检验对参数进行特征筛选。基于二元Logistic回归构建血液学模型、CT纹理分析模型及综合模型来预测VEGFR2表达状态。通过受试者工作特征曲线评估三个模型的诊断效能,并通过列线图来可视化地预测胃癌患者VEGFR2的表达状态。结果基于血液学指标构建的血液学模型曲线下面积(area under the curve,AUC)为0.687。由静脉期纹理分析参数构建的CT纹理分析模型的AUC值为0.624。联合血液学模型和CT纹理分析模型构建的综合模型AUC值为0.723。结论联合多期动态增强CT纹理分析参数及血液学指标有助于术前预测胃癌VEGFR2表达状态。

白屈菜红碱对人卵巢癌SKOV3细胞迁移、侵袭和上皮-间质转化的影响

目的:探讨白屈菜红碱(CHE)对人卵巢癌SKOV3细胞迁移、侵袭和上皮-间质转化(EMT)的抑制作用,阐明其相关作用机制。方法:体外培养SKOV3细胞,分为对照组和2.5、5.0、10.0、20.0及40.0μmol·L^(-1)CHE组,采用噻唑蓝(MTT)法检测各组细胞增殖抑制率。体外培养SKOV3细胞,分为对照组、转移生长因子β1(TGF-β1)组、TGF-β1+5μmol·L^(-1)CHE组和TGF-β1+10μmol·L^(-1)CHE组,采用细胞划痕实验检测各组细胞迁移率,Transwell小室实验检测各组细胞中迁移细胞数和侵袭细胞数,Westren blotting法检测各组细胞中E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)蛋白表达水平,免疫荧光染色法检测各组细胞中E-cadherin和N-cadherin荧光强度。结果:MTT法,与对照组比较,5.0、10.0、20.0和40.0μmol·L^(-1)CHE组细胞增殖抑制率明显升高(P<0.05或P<0.01)。细胞划痕实验,与对照组比较,TGF-β1组细胞迁移率明显升高(P<0.01);与TGF-β1组比较,TGF-β1+5μmol·L^(-1)CHE组和TGF-β1+10μmol·L^(-1)CHE组细胞迁移率明显降低(P<0.01)。Transwell小室实验,与对照组比较,TGF-β1组细胞中迁移细胞数和侵袭细胞数明显增加(P<0.05);与TGF-β1组比较,TGF-β1+5μmol·L^(-1)CHE组和TGF-β1+10μmol·L^(-1)CHE组细胞中迁移细胞数和侵袭细胞数明显减少(P<0.01)。Westren blotting法,与对照组比较,TGF-β1组细胞中E-cadherin蛋白表达水平明显降低(P<0.01),N-cadherin和Vimentin蛋白表达水平明显升高(P<0.05或P<0.01);与TGF-β1组比较,TGF-β1+5μmol·L^(-1)CHE组和TGF-β1+10μmol·L^(-1)CHE组细胞中E-cadherin蛋白表达水平明显升高(P<0.01),N-cadherin和Vimentin蛋白表达水平明显降低(P<0.01)。免疫荧光染色,与对照组比较,TGF-β1组细胞中E-cadherin荧光强度明显降低,N-cadherin荧光强度明显升高;与TGF-β1组比较,TGF-β1+5μmol·L^(-1)CHE组和TGF-β1+10μmol·L^(-1)CHE组细胞中E-cadherin荧光强度明显升高,N-cadherin荧光强度明显降低。结论:CHE能够抑制人卵巢癌SKOV3细胞增殖、迁移、侵袭和EMT。

微RNA-196a-1-3p靶向Ras响应元件结合蛋白调控胆管癌细胞增殖的机制研究

目的探讨转化生长因子β(TGF-β)调控人胆管癌细胞系RBE细胞增殖的关键微RNA(miRNA)及其潜在的机制。方法该研究起止时间为2020年1月至2022年1月。磷酸盐缓冲液(PBS)处理为对照组,TGF-β处理为TGF-β组,TGF-β抗体处理为抗体组。检测三组RBE细胞的增殖水平。miRNA高通量测序检测三组RBE细胞的miRNA调控变化,并进行miRNA模拟物过表达筛选鉴定受TGF-β调控的影响RBE细胞增殖水平的关键miRNA。miRNA数据库(miRDB)在线分析miRNA的潜在底物,并通过小干扰RNA(siRNA)敲低筛选鉴定影响RBE细胞增殖水平的关键底物。结果相比于对照组,TGF-β组RBE细胞的增殖水平上升(1.62±0.07比2.35±0.09,P<0.05),抗体组RBE细胞的增殖水平下降(1.62±0.07比1.11±0.08,P<0.05)。过表达微RNA-196a-1-3p(miR-196a-1-3p)时,RBE细胞的增殖水平下降(P<0.05)。敲低Ras响应元件结合蛋白(RREB1)时,RBE细胞的增殖水平下降(P<0.05)。过表达miR-196a-1-3p后,RBE细胞中RREB1的信使RNA(mRNA)和蛋白水平下降(P<0.05)。敲低miR-196a-1-3p后,RBE细胞中RREB1与SMAD家族蛋白3(SMAD3)的相互作用增加。敲低SMAD3后,RBE细胞的增殖水平下降(P<0.05)。与仅敲低SMAD3相比,敲低SMAD3的同时过表达RREB1的RBE细胞的增殖水平无显著变化,并且同时敲低SMAD3和miR-196a-1-3p的RBE细胞的增殖水平无显著变化。结论TGF-β能够通过miR-196a-1-3p/RREB1/SMAD3轴促进RBE细胞增殖;miR-196a-1-3p和RREB1可作为潜在的治疗胆管癌的靶标,为针对该靶标的新药研发奠定了基础。

TGF-β对膀胱癌组织中巨噬细胞来源肿瘤相关成纤维细胞的影响及其机制

目的探讨TGF-β对膀胱癌组织中巨噬细胞来源肿瘤相关成纤维细胞(CAF)的影响及其机制。方法采用Kaplan-Meier法分析膀胱癌组织中α-SMA^(+)CD68^(+)CAF表达水平与患者的总生存率(OS)的关系;采用免疫荧光技术检测α-SMA^(+)CD68^(+)CAF在膀胱癌组织中的浸润情况;采用Western blot实验检测TGF-β对α-SMA^(+)CD68^(+)CAF体外诱导作用。同时构建膀胱癌小鼠模型,采用免疫荧光技术和免疫组化法检测TGF-β对膀胱癌小鼠体内α-SMA^(+)CD68^(+)CAF的诱导作用及对CD8^(+)T细胞浸润的影响。结果膀胱癌组织中α-SMA与CD68的表达呈正相关,且α-SMA^(+)CD68^(+)CAF高表达的患者预后更差(χ^(2)=9.05,P<0.05)。免疫荧光技术检测结果显示,膀胱癌组织中存在α-SMA^(+)CD68^(+)CAF。Western blot实验检测结果显示,TGF-β可显著促进α-SMA^(+)CD68^(+)CAF的生成。膀胱癌小鼠模型体内实验显示,TGF-β可促进膀胱癌组织中α-SMA^(+)CD68^(+)CAF的生成,从而抑制CD8^(+)T细胞的浸润。结论TGF-β可显著促进膀胱癌组织中巨噬细胞来源CAF的生成,从而抑制CD8^(+)T细胞的浸润。

miR-484通过调控HIF-1α/VEGF信号通路对胃癌血管生成的影响

【目的】探讨miR-484是否通过调控HIF-1α/VEGF信号通路参与胃癌血管的生成。【方法】分别在阴性对照(miR-NC组)、miR-484模拟物(miR-484 mimics组)和miR-484抑制剂(miR-484 inhibitors组)3种条件下转染SCG-7901胃癌细胞。检测不同组SCG-7901细胞中缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)的mRNA和蛋白表达情况及胃癌细胞的增殖情况。将转染成功的SCG-7901细胞与血管内皮细胞分别在缺氧条件下共培养,采用ELISA法检测培养基中HIF-1α和VEGF的表达情况,CCK8检测血管内皮细胞的增殖情况。【结果】miR-484 mimics处理后,SCG-7901细胞的增殖能力显著降低,而miR-484 inhibitor处理组的细胞增殖能力则显著提高。与miR-NC组比较,miR-484 mimics组SCG-7901细胞的HIF-1α、VEGF mRNA和蛋白表达下调(均P<0.01),miR-484 inhibitor组的结果与之相反。在缺氧条件下,与miR-NC组相比,miR-484 mimics显著降低了培养基中HIF-1α和VEGF水平(均P<0.05),血管内皮细胞增殖能力降低(P<0.05),而miR-484 inhibitor组的结果与之相反。【结论】miR-484可能通过下调HIF-1α/VEGF通路抑制胃癌细胞增殖和肿瘤血管形成。

抗体药物偶联物在人表皮生长因子受体2低表达胃癌中的应用研究进展

胃癌(GC)是极具异质性和侵袭性的消化系统恶性肿瘤之一,传统化疗药物及曲妥珠单抗等人表皮生长因子受体2(HER2)靶向药物在GC的治疗过程中仍然存在耐药性发生率高、毒副作用大、患者耐受差等缺点。因此,研发更为有效的抗GC药物势在必行。目前针对HER2的新型靶向药层出不穷,但在某些情况下无效或产生耐药,这与HER2在某些GC细胞中低表达有关,HER2低表达(HER2 IHC1+或IHC2+/ISH-)约占全部类型的40%~60%,但在临床实践中,这类患者仍被报告为HER2阴性GC。因此准确检测HER2表达状态对于确定可能受益于曲妥珠单抗治疗的患者至关重要。抗体药物偶联物(ADC)的出现为HER2阳性GC提供了新的治疗选择,凭借其精准高效的抗肿瘤作用,有望在未来替代传统GC化学疗法。近期有研究发现ADC可能在HER2低表达GC中具有潜在抗肿瘤活性,相关临床研究正在评估其在HER2低表达GC治疗中的有效性和安全性。本文就靶向治疗时代ADC在HER2低表达GC患者中的应用和最新研究进展作一综述,并讨论HER2靶向ADC在应用和研发过程中面临的挑战。

Inhibitory effects of RRR-α-tocopheryl succinate on benzo (a) pyrene (B (a) P)-induced forestomach carcinogenesis in female mice

AIM To study the inhibitory effects of VES( RRR-α-tocopheryl Succinate, VES ), aderivative of natural Vitamin E, on benzo (a)pyrene (B (a) P)-induced forestomach tumor infemale mice.METHODS The model of B (a)P-inducedforestomach tumor was established according tothe methods of Wattenberg with slightmodifications. One hundred and eighty femalemice (6 weeks old) were divided into six groupsequally; negative control (Succinic acid),vehicle control ( Succinate + B (a) P), positivecontrol(B(a) P), high VES(2.5g/kg. b. w + B(a)P), Iow VES(1 .25 g/kg. b. w + B(a) P) ig as wellas VES by ip (20 mg/kg, b. w + B(a) P). Exceptthe negative control group, the mice wereadministrated with B(a)P ig. and correspondingtreatments for 4 weeks to study the anti-carcinogenetic effect of VES during the initiationperiod. The experiment lasted 29 weeks, inwhich the inhibitory effects of VES both ontumor incidence and tumor size were tested.RESULTS The models of B (a)P-inducedforestomach tumor in female mice wereestablished successfully. Some werecauliflower-like, others looked like papilla, evena few were formed into the ulcer cavities. VES at1.25 g/kg. b. w, 2.5 g/kg. b.w. by ig and 20 mg/kg. b. w. via ip could decrease the number oftumors per mouse (1.7 ± 0. 41, 1.6 ± 0.34 and 1.1±0.43), being lower than that of B(a)P group(5.4 ± 0.32, P＜0.05). The tumor incidence wasinhibited by 18.2%, 23.1% and 50.0%. VES at1.25g/kg.b.w., 2.5 g/ kg.b.w. by ig and20 mg/kg. b.w. via ip reduced the total volumeof tumors per mouse (54.8 ± 8.84, 28.4 ± 8.32and 23.9± 16.05), being significantly lower thanthat of B(a)P group (150.2±20.93, P＜0.01).The inhibitory rates were 63.5%, 81.1% and84.1%, respectively.CONCLUSION VES has inhibitory effects on B(a) P-induced forestomach carcinogenesis infemale mice, especially by ip and it may be apotential anti-cancer agent in vivo.

Clinicopathologic factors and molecular markers related to lymph node metastasis in early gastric cancer

AIM: To analyze predictive factors for lymph node metastasis in early gastric cancer.METHODS: We analyzed 1104 patients with early gastric cancer(EGC) who underwent a gastrectomy with lymph-node dissection from May 2003 through July 2011. The clinicopathologic factors and molecular markers were assessed as predictors for lymph node metastasis. Molecular markers such as microsatellite instability, human mut L homolog 1, p53, epidermal growth factor receptor(EGFR) and human epidermal growth factor receptor 2(HER2) were included. The χ2 test and logistic regression analysis were used to determine clinicopathologic parameters.RESULTS: Lymph node metastasis was observed in 104(9.4%) of 1104 patients. Among 104 cases of lymph node positive patients, 24 patients(3.8%) were mucosal cancers and 80 patients(16.7%) were submucosal. According to histologic evaluation, the number of lymph node metastasis found was 4(1.7%) for well differentiated tubular adenocarcinoma, 45(11.3%) for moderately differentiated tubular adenocarcinoma, 36(14.8%) for poorly differentiated tubular adenocarcinoma, and 19(8.4%) for signet ring cell carcinoma. Of 690 EGC cases, 77 cases(11.2%) showed EGFR overexpression. HER2 overexpression was present in 110 cases(27.1%) of 406 EGC patients. With multivariate analysis, female gender(OR = 2.281, P = 0.009), presence of lymphovascular invasion(OR = 10.950, P < 0.0001), diameter(≥ 20 mm, OR = 3.173, P = 0.01), and EGFR overexpression(OR = 2.185, P = 0.044) were independent risk factors for lymph node involvement.CONCLUSION: Female gender, tumor size, lymphovascular invasion and EGFR overexpression were predictive risk factors for lymph node metastasis in EGC.