Beclin 1 is involved in autophagy, differentiation, apoptosis and cancer progression, and functions as a haploinsufficient tumor suppressor gene. The aim of the present study was to elucidate the function of Beclin 1 in colon cancer. A Beclin 1-expressing plasmid was transfected into HCT-15 and HCT-116 cells, and the phenotypes and associated molecules were determined. Beclin 1 transfectants were subcutaneously injected into nude mice to determine tumor growth, and proliferation and apoptosis levels using Ki-67 immunostaining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), respectively. Beclin 1 overexpression inhibited viability as determined using a Cell Counting Kit-8 assay, inhibited migration and invasion as determined using a wound healing assay or Transwell assay, and lamellipodia formation by filamentous actin staining, induced autophagy as determined using electron microscopy, and light chain 3B (LC-3B) expression, and apoptosis as determined using Annexin V staining in the two cell lines (P<0.05). Beclin 1 induced G2 arrest of HCT-15 transfectants as determined using propidium iodide staining (P<0.05), whereas HCT-116 transfectants were arrested in G1 phase (P<0.05). The two transfectants exhibited increased expression of c-Myc, cyclin D1, β-catenin, insulin-response element 1 and 78 kDa glucose-regulated protein compared with the control and mock cells as determined using the reverse transcription-quantitative polymerase chain reaction (P<0.05). Beclin 1 overexpression upregulated LC-3B and cyclin-dependent kinase 4 expression, but downregulated cyclin E expression of the cancer cell lines as determined using western blot analysis (P<0.05). Beclin 1 expression in vivo significantly suppressed the proliferation of colon cancer cells in xenograft models via inducing apoptosis by TUNEL, and inhibiting proliferation by Ki-67 expression (P<0.05). Beclin 1 overexpression may reverse aggressive phenotypes and suppress colon cancer tumor growth, and be employed as a target molecule for gene therapy of patients with colon cancer.

In the present study, SGC-7901/DDP cells were treated with different concentrations of parthenolide (PN) (2.5-15 µmol/l), cisplatin (DDP) (1.25-15 µg/ml) and PN+DDP. The proliferation inhibition rates were measured using an MTT assay, and the synergies of PN and DDP were analyzed. The effect of PN and DDP on SGC-7901/DDP cell proliferation demonstrated a time- and concentration-dependent association, and a synergy between PN and DDP was identified. DAPI staining and flow cytometry results indicated that 15 µmol/l PN significantly induced SGC-7901/DDP apoptosis and G1 phase arrest compared with the untreated control group. Western blotting analysis results indicated that among the apoptosis-associated proteins, there were dose-dependent increases in the protein expression of apoptosis regulator BAX, cellular tumor antigen p53, cleaved caspase-3 and cleaved capase-9, and decreases in apoptosis regulator Bcl-2 and Bcl-xL protein expression levels. Among the cell cycle-associated proteins, cyclin D1 expression was significantly decreased, cyclin-dependent kinase inhibitor 1 expression was significantly increased, and signal transducer and activator of transcription 3 (STAT3) activation was inhibited. Scratch and Transwell assay results revealed that PN significantly inhibited SGC-7901/DDP cell migration, and invasion. The present study demonstrated that PN induces SGC-7901/DDP apoptosis, inhibits SGC-7901/DDP proliferation, migration and invasion, and enhances the drug sensitivity of the cells to DDP. The underlying mechanisms may be associated with inhibition of the STAT3 signaling pathway and regulation of the downstream apoptotic protein and cyclin expression levels.

Here, we found that both SAHA and MG132 synergistically inhibited proliferation, glycolysis and mitochondrial oxidization, induced cell cycle arrest and apoptosis in MGC-803 and MKN28 cells. SAHA increased cell migration and invasionat a low concentration. SAHA induced the overexpression of acetyl histone 3 and 4, which were recruited to p21, p27, Cyclin D1, c-myc and nanog promoters to transcriptionally up-regulate the former two and down-regulate the latter three. The expression of acetyl-histone 3 and 4 was increased during gastric carcinogenesis and positively correlated with cancer differentiation. SAHA and MG132 exposure suppressed tumor growth by inhibiting proliferation and inducing apoptosis in nude mice, increased serum ALT and AST levels and decreased hemaglobin level, white blood cell and neutrophil numbers. These data indicated that SAHA and MG132 in vivo and vitro synergistically induced cytotoxicity and apoptosis, suppressed proliferation, growth, migration and invasion of gastric cancer cells. Therefore, they might potentially be employed as chemotherapeutic agents if the hepatic injury and the killing effects of peripheral blood cells are avoided or ameliorated.

Inhibitor of growth protein 2 (ING2) has an important role in the regulation of chromatin remodeling, cell proliferation, cell‑cycle arrest, senescence and apoptosis. The present study performed an immunohistochemical analysis for expression profiling of ING2 protein in an array of tissues comprising normal mouse and human tissues, as well as human hepatocellular (n=62), renal clear cell (n=62), pancreatic (n=62), esophageal squamous cell (n=45), cervical squamous cell (n=31), breast (n=144), gastric (n=196), colorectal (n=96), ovarian (n=208), endometrial (n=96) and lung (n=192) carcinoma tissues. In mouse tissues, ING2 was detected in the nuclei and cytoplasm of the glandular epithelium of breast, hepatocytes, intestine, bronchium and alveoli, as well as the squamous epithelium of skin and glomeruli, and in myocardial cells, while it was located in the cytoplasm of renal tubules and striated muscle cells. ING2 protein was scattered in the brain and spleen. In human tissues, ING2 protein was principally distributed in the cytoplasm, while in it was present in the cytoplasm and nuclei in the stomach, intestine, cervix, endometrium trachea, breast and pancreas. The nuclear location of ING2 in the stomach was more prominent than that in the cytoplasm. High ING2 immunoreactivity was detected in the tongue, stomach, skin, pancreas, cervix and breast, whereas weakly in the brain stem, thymus, thyroid, lung, striated muscle, testis, bladder and ovary. In total, 617 out of 1,194 of the tested cancer tissues (51.7%) were ING2-positive. In most cases, ING2 expression was found to be restricted to the cytoplasm of all cancer tissues, while in certain cancer types, including renal clear cell, ovarian and colorectal carcinoma, it was occasionally present in the nuclei. Among the cancer tissues examined, ING2 was most frequently expressed in breast cancer (67.4%) and gynecological cancer types, including ovarian cancer (61.5%) and endometrial cancer (57.3%). Compared with that in the respective normal tissues, ING2 expression in breast cancer tissues was decreased, while that in cervical cancer was upregulated in the nuclei as well as the cytoplasm. In endometrial cancer, expression of ING2 was increased in the nuclei and declined in the cytoplasm compared with that in the normal endometrium. ING2‑positive cases were less frequent for renal clear cell carcinoma (17.7%). The results of the present study suggested that ING2 may be involved in the repair and regeneration of organs or tissues and is associated with breast and gynecological carcinogenesis.

目的:探讨慢病毒介导的CXCR7-shRNA转染人结肠癌细胞株SW620后对CXCR7蛋白表达的影响。方法:（1）设计并合成CXCR7的3对shRNA序列及1对阴性对照序列,与pSilencerTM4.1系统合成构建重组慢病毒载体,转染HEK293T细胞包装病毒并检测滴度;（2）将3种重组慢病毒载体及阴性对照分别感染人结肠癌细胞SW620,RT-PCR检测CXCR7 mRNA的表达情况,测定沉默效率,筛选沉默效率最高的一组CXCR7-shRNA作为后续实验表达载体;（3）MTT法检测转染CXCR7-shRNA对SW620细胞生长增殖的影响;（4）通过细胞划痕实验检测转染CXCR7-shRNA对SW620细胞侵袭迁移能力的影响;（5）Western blot检测转染CXCR7-shRNA对SW620细胞蛋白表达情况。结果:（1）测序证实3对慢病毒载体及1对阴性对照载体均包装成功,滴度分别为3.16×108TU/ml、4.27×108TU/ml、3.93×108TU/ml和2.95×108TU/ml;（2）3组慢病毒载体转染SW620细胞后,CXCR7 mRNA的表达量均较阴性对照组明显降低（P＜0.05）,其中CXCR7-shRNA-1对CXCR7的抑制率明显高于其他两组（P＜0.05）;（3）CXCR7-shRNA-1转染SW620后,肿瘤细胞的增殖程度显著减少,与空白组相比有显著性差异（P＜0.05）;（4）SW620细胞在划痕24h后,空白对照组和实验组的细胞迁移指数（MI）分别为（49.92±6.41）%和（29.13±5.38）%,有统计学意义（P＜0.05）,划痕48h后,对照组与实验组的MI分别为（96.52±7.44）%和（72.03±8.29）%,有统计学意义（P＜0.05）;（5）CXCR7-shRNA-1转染SW620细胞后,与空病毒载体组、空白组相比,CXCR7蛋白表达量明显降低,具有统计学意义（P＜0.05）。结论:CXCR7-shRNA慢病毒表达载体转染SW620细胞后可有效下调CXCR7 mRNA和蛋白的表达水平并能够抑制肿瘤细胞的增殖与迁移,为下一步研究以CXCR7/CXCL12生物学轴为靶点的结直肠癌慢病毒基因沉默治疗打下了良好的基础,为结直肠癌的基因治疗提供了新方向。

目的 探讨慢病毒介导的CXCR7-shRNA转染人结肠癌细胞株SW620后对CXCR7蛋白表达的影响.方法 ①设计并合成CXCR7的3对shRNA序列及1对阴性对照序列,与pSilencerTM4.1系统合成构建重组慢病毒载体,转染HEK293T细胞包装病毒并检测滴度;②将3种重组慢病毒载体及阴性对照分别感染人结肠癌细胞SW620,RT-PCR检测CXCR7 mRNA的表达情况,测定沉默效率,筛选沉默效率最高的一组CXCR7-shRNA作为后续实验的表达载体;③MTT法检测转染CXCR7-shRNA对SW620细胞生长增殖的影响;④细胞划痕实验检测转染CXCR7shRNA对SW620细胞侵袭迁移能力的影响;⑤Western blot检测转染CXCR7-shRNA对SW620细胞蛋白表达情况.结果 ①测序证实3对慢病毒载体及1对阴性对照载体均包装成功,滴度分别3.16×108 pfu/mL、4.27×108 pfu/mL、3.93×108 pfu/mL和2.95×108 pfu/mL;②3组慢病毒载体转染SW620细胞后,CXCR7 mRNA的表达量均较阴性对照组明显降低(P＜0.05),其中CXCR7-shRNA-1对CXCR7的抑制率明显高于其他两组(P＜0.05);③CXCR7-shR-NA-1转染SW620后,肿瘤细胞的增殖程度显著减少,与空白组相比有统计学差异(P＜0.05);④SW620细胞在划痕24 h后,空白对照组和实验组的细胞迁移指数(MI)分别为(49.92±6.41)％和(29.13±5.38)％,差异有统计学意义(P＜0.05),划痕48h后,对照组与实验组的MI分别为(96.52±7.44)％和(72.03±8.29)％,差异有统计学意义(P＜0.05);⑤CXCR7-shRNA-1转染SW620细胞后,与空病毒载体组、空白组相比,CXCR7蛋白表达量明显降低,具有统计学意义(P＜0.05).结论 CXCR7被沉默后可有效抑制SW620肿瘤细胞的增殖与迁移,这为后续研究以CXCR7/CXCL12生物学轴为靶点的肿瘤基因治疗打下良好基础.

Objective To investigate the CXCR 7 protein expression when CXCR 7-shRNA transfected into human gastric cancer cell which mediated with lentivirus vector combined with Rhizoma Paridis Total Saponin. Methods Three shRNA sequences of CXCR 7 and one negative control sequence were designed and synthesized, and recombinant lentiviral vectors with pSilencerTM 4.1 system were established. Transfection of HEK 293 T cells and packaging viral were finished and the titers were detected. Transfection of all recombinant lentiviral vectors and negative control vector were finished and expression of CXCR 7 mRNA were detected by RT-PCR method. Silence efficiency in groups were determined and the expression vector with highest silence efficiency was selected for next experiments. To detect the effect of SGC 7901 cell proliferation by CXCR 7-shRNA transfection and combined with Rhizoma Paridis Total Saponin intervention with MTT. To detect the effect of SGC 7901 cell expression of protein by CXCR 7-shRNA transfection and combined with Rhizoma Paridis Total Saponin intervention with Western blot. Results The packaging of three lentiviral vector and negative control sequence are successful which is confirmed by gene sequencing and the titer are 4.9×108 pfu/mL, 3.6×108 pfu/mL, 5.2×108 pfu/mL, 2.0×108 pfu/mL respective. The expression quantity of CXCR 7 mRNA in positive groups are lower than negative control group(P<0.05)and inhibition ratio to CXCR 7 in CXCR 7-shRNA-1 and combined with Rhizoma Paridis Total Saponin intervention group is higher than the other two groups(P<0.05). The proliferation level of tumour cell is significant reduction after CXCR 7-shRNA-1 transfection and have a significant difference comparing to the group without transfection(P<0.05). The expression of CXCR 7 protein is significant reduction after CXCR 7-shRNA-1 transfection comparing to the group without virus vector and negative control group and have a significant difference(P<0.05). Conclusion The construction of three CXCR 7-shRNA lentiviral expression vector are successful and expression level of protein and CXCR 7 mRNA are down-regulated effectively after transfection and combined with Rhizoma Paridis Total Saponin intervention. It maybe means that CXCR 7 gene takes an important role in the process of gastric cancer proliferation and invasion.This is foundation for further study of gastric cancer gene therapy using CXCR 7/CXCL 12 biological axis as a target.

Background RhoC is a small G protein/GTPase and involved in tumor mobility, invasion and metastasis. Previously, up-regulated RhoC expression is found to play an important role in ovarian carcinogenesis and subsequent progression by modulating proliferation, apoptosis, migration and invasion. Methods We transfected RhoC-expressing plasmid and RhoC siRNA into CAOV3 and OVCAR3 cells respectively. These cells and transfectants were exposed to vascular epithelial growth factor (VEGF), transforming growth factor (TGF)-β1 or their receptor inhibitors with the phenotypes and their related-molecules examined. Results TGF-β1R or VEGFR inhibitor suppressed the proliferation, migration, invasion and lamellipodia formation, the expression of N-cadherin, α-SMA, snail and Notch1 mRNA or protein, and enhanced E-cadherin mRNA and protein expression in CAOV3 and its RhoC-overexpressing transfectants, whereas both growth factors had the opposite effects in OVCAR3 cells and their RhoC-hypoexpressing transfectants. Ectopic RhoC expression enhanced migration, invasion, lamellipodia formation and the alteration in epithelial to mesenchymal transition (EMT) markers of CAOV3 cells regardless of the treatment of VEGFR or TGF-β1R inhibitor, whereas RhoC knockdown resulted in the converse in OVCAR3 cells even with the exposure to VEGF or TGF-β1. Conclusion RhoC expression might be involved in EMT of ovarian epithelial carcinoma cells, stimulated by TGF-β1 and VEGF.

The molecular mechanism underlying the proliferation of colorectal cancer (CRC) cells is not completely understood. Here, we found that the level of neuregulin receptor degradation protein-1 (Nrdp1) E3 ubiquitin ligase was significantly decreased in CRC tissues, compared with the adjacent normal tissues from human patients. Knockdown of Nrdp1 enhanced the proliferation of CRC cells, while overexpression of Nrdp1 inhibited the proliferation of CRC cells. Further analysis showed that Nrdp1 may induce degradation of its target ErbB3 to inhibit activation of both ERK/MAPK and PI3K/Akt pathways in CRC cells, which seemed to affect cell proliferation via nuclear retention of a major cell-cycle inhibitor, p27. Taken together, these findings suggest that Nrdp1-mediated ErbB3 degradation suppresses cellular growth of CRC and that Nrdp1 loss in CRC may promote tumor progression, thus highlighting Nrdp1 as a novel target for CRC therapy.

Neovascularization plays a substantial role in the invasiveness and metastasis of gastric cancer. However, the molecular mechanism underlying the control of neovascularization of gastric cancer remains undefined. Both vascular endothelial growth factor a (VEGFa) and matrix metalloproteinases 2 (MMP2) are essential for neovascularization by promoting endothelial mitogenesis and permeability and by promoting extracellular matrix degradation, respectively. Therefore, we were prompted to examine whether VEGFa and MMP2 may affect expression of each other to coordinate the neovascularization process. We found strong positive correlation of VEGFa and MMP2 levels in the gastric patients. Moreover, patients with metastasis of the original cancer had significantly higher levels of VEGFa and MMP2. Thus, we used a human gastric cancer cell line, SNU-5, to examine whether expression of VEGFa and MMP2 may affect each other. We found that overexpression of VEGFa in SNU-5 cells increased expression of MMP2, while inhibition of VEGFa in SNU-5 cells decreased expression of MMP2. On the other hand, overexpression of MMP2 in SNU-5 cells increased expression of VEGFa, while inhibition of MMP2 in SNU-5 cells decreased expression of VEGFa. These data suggest that expression of VEGFa and MMP2 may activate each other to reinforce a promoting effect on neovascularization. We then analyzed how VEGFa expression affects MMP2 level. Application of a specific Akt inhibitor to VEGFa-overexpressing SNU-5 cells substantially abolished the effect of VEGFa on MMP2 activation, suggesting that VEGFa may increase expression of MMP2 via PI3K/Akt signaling pathway. Since anti-VEGFa is a well-established therapy for many cancers, our data suggest that anti-VEGFa may not only inhibit neovascularization by prohibiting VEGFa-dependent endothelial mitogenesis and permeability increase but also by downregulating MMP2 to abolish the extracellular matrix degradation in gastric cancer.

Isoliquiritigenin, a herbal ingredient with chalcone structure, has been speculated to be able to inhibit one of the most drug-metabolizing enzymes (DMEs) UDP-glucuronosyltransferase (UGT). Therefore, the aim of the present study was to investigate the inhibition of isoliquiritigenin towards important UGT isoforms in the liver and intestine, including UGT1A1, 1A3, 1A6, 1A7, 1A8, 1A9 and 1A10. The recombinant UGT-catalyzed 4-methylumbelliferone (4-MU) glucuronidation was used as probe reactions. The results showed that 100μM of isoliquiritigenin inhibited the activity of UGT1A1, UGT1A3, UGT1A6, UGT1A7, UGT1A8, UGT1A9, and UGT1A10 by 95.2%, 76.1%, 78.9%, 87.2%, 67.2%, 94.8%, and 91.7%, respectively. The data fitting using Dixon plot and Lineweaver-Burk plot showed that the inhibition of UGT1A1, UGT1A9 and UGT1A10 by isoliquiritigenin was all best fit to the competitive inhibition, and the second plot using the slopes from the Lineweaver-Burk plot versus isoliquiritigenin concentrations was used to calculate the inhibition kinetic parameter (K(i)) to be 0.7μM, 0.3μM, and 18.3μM for UGT1A1, UGT1A9, and UGT1A10, respectively. All these results indicated the risk of clinical application of isoliquiritigenin on the drug-drug interaction and other possible diseases induced by the inhibition of isoliquiritigenin towards these UGT isoforms.

Objective To investigate the inhibitory effect of the CXCR4-shRNA adenovirus vector on the growth and metastasis of transplanted tumors of human gastric cancer cells in nude mice.Methods Three groups of nude mice tumor model with abdominal cavity metastasis were established:the SGC7901 group,the vector virus group,the CXCR4-shRNA group.The diameter,weight of the tumor and the number of metastatic tumor in the three groups of nude mice tumor were compared.Their survival time,survival rate,life extension rate were compared.Western blot was used to detect the VEGF-C and MMP-2 protein expression levels.Results Compared with the SGC7901 group and the vector virus group,the ascites production time in CXCR4-shRNA group nude mice was significantly delayed and less,the diameter and weight of tumor were decreased,as well as the survival time was extended.In addition,the mortality was reduced in the nude mice.The protein expression levels of the VEGF-C,MMP-2 were inhibited.Conclusion CXCR4-shRNA adenovirus vector can inhibit growth and metastasis of human gastric carcinoma SGC7901 cell in nude mice.

BACKGROUND Parafibromin is a protein encoded by the HRPT2 (hyperparathyroidism 2) oncosuppressor gene and its down-regulated expression is involved in pathogenesis of parathyroid, breast, gastric and colorectal carcinomas. This study aimed to clarify the effects of parafibromin expression on the phenotypes and relevant mechanisms of DLD-1 colon carcinoma cells. METHODS DLD-1 cells transfected with a parafibromin-expressing plasmid were subjected to examination of phenotype, including proliferation, differentiation, apoptosis, migration and invasion. Phenotype-related proteins were measured by Western blot. Parafibromin and ki-67 expression was detected by immunohistochemistry on tissue microarrays. RESULTS The transfectants showed higher proliferation by CCK-8, better differentiation by electron microscopy and ALP activity and more apoptotic resistance to cisplatin by DNA fragmentation than controls. There was no difference in early apoptosis by annexin V, capase-3 activity, migration and invasion between DLD-1 cells and their transfectants. Ectopic parafibromin expression resulted in down-regulated expression of smad4, MEKK, GRP94, GRP78, GSK3β-ser9, and Caspase-9. However, no difference was detectable in caspase-12 and -8 expression. A positive relationship was noted between parafibromin and ki-67 expression in colorectal carcinoma. CONCLUSIONS Parafibromin overexpression could promote cell proliferation, apoptotic resistance, and differentiation of DLD-1 cells.

The polymorphism of X-ray repair cross-complementing group 1 (XRCC1) Arg194Trp, a substitution of Arg to Gln at position 194, has been implicated in the development of colorectal cancer (CRC) in a number of case–control studies with contradictory and inconclusive findings. The current meta-analysis of all currently available publications was conducted to assess the gene susceptibility to CRC and improve our understanding of the CRC pathogenesis. The pooled odds ratio (OR) with corresponding 95 % confidence interval (95 % CI) was calculated by use of fixed-effects model or random-effects model when appropriate. A total of 15 eligible case–control studies with 4,501 cases and 8,038 controls were retrieved after a comprehensive search of the PubMed, Embase, Web of science, and Chinese Biomedicine (CBM) databases up to December 2012. The overall meta-analysis identified a positive but not statistically significant association between the XRCC1 Arg194Trp polymorphism and CRC risk under all genetic contrast models (OR Trp vs. Arg = 1.07, 95 % CI 0.90–1.26, P OR = 0.441; OR TrpTrp vs. ArgArg = 1.28, 95 % CI 0.91–1.81, P OR = 0.163; OR ArgTrp vs. ArgArg = 1.00, 95 % CI 0.85–1.19, P OR = 0.956; OR ArgTrp + TrpTrp vs. ArgArg = 1.06, 95 % CI 0.90–1.24, P OR = 0.502; OR TrpTrp vs. ArgArg + ArgTrp = 1.11, 95 % CI 0.91–1.34, P OR = 0.306). The genotype TrpTrp carriers among Caucasians were more susceptible to CRC, although lack statistical evidence (OR TrpTrp vs. ArgArg = 2.69, 95 % CI 0.97–7.49, P OR = 0.058; OR TrpTrp vs. ArgArg + ArgTrp = 2.77, 95 % CI 0.99–7.72, P OR = 0.051). Interestingly, the XRCC1 Arg194Trp variant was significantly associated with an increased risk of colon cancer. The present meta-analysis suggests that the XRCC1 Arg194Trp polymorphism may modify the risk for CRC, particularly colon cancer. However, the precise genetic association needs to be further estimated in future studies.

目的探讨特异性沉默胃癌细胞SCG-7901中的趋化因子受体4(CXCR4)的检验效率及其进行动物实验的可行性。方法将CX-CR4的特异性RNA干扰载体转染至胃癌细胞SCG-7901 72 h后,倒置荧光显微镜观察转染情况,测定最优的MOI值,采用RT-PCR和Western印迹法进行效率测定。结果 (1)成功将CXCR4-shRNA重组腺病毒载体转染至胃癌细胞SGC7901。(2)空白组与对照组CXCR4 mRNA及蛋白表达量明显高于实验组,有统计学意义(P<0.01),而空白组与对照组之间无显著性差异(P>0.05)。(3)沉默效率为82.7%。结论重组腺病毒载体CXCR4-shRNA可抑制胃癌细胞SCG7901中CXCR4基因mRNA和蛋白的表达,并可作为动物实验的理论基础和前期条件。

Objective To detect the influence of RNA interference fragment to ability of cell proliferation in SGC7901 cells with specific inhibition of NRP2 gene expression in gastric cancer cell line namely SGC7901.Methods NRP2 adenovirus vector was transfered into SCG7901 cells.After 72 hours,SCG7901 cells were observed.expression of mRNA in NRP2 gene was detected by RT-PCR,proliferation of gastric cancer cell was detected by MTT and protein expression of NRP2 was detected after transfection by Western blot respectively.Results The transfection of NRP2-shRNA recombinant adenovirus vector into gastric cancer cell SGC7901 was successful.Level of NRP2 mRNA in transfection group was lower than that of blank and negative control groups and inhibitory effect of NRP2-shRNA-3 was the strongest.The cell proliferation was more rapidly in blank and negative control groups and there was no significant difference(P>0.05).The cell proliferation was depressed in transfection group and there was significant difference compared to blank group or negative control group(P<0.05).Expression content in transfection group was higher than that of another two groups and there was significant difference(P<0.05) but there was no significant difference between blank and negative control groups(P>0.05).Conclusions The expression of mRNA and protein of NRP2 in gastric cancer cell SGC7901 will be inhibited by NRP2-shRNA recombinant adenovirus vector and the same influence to cancer cell in some degree.It is theoretical basis and prophase conditions for animal experiment.

BACKGROUND:Oncogenic Bmi-1 (B-lymphoma Moloney murine leukemia virus insertion region-1) belongs to the Polycomb-group (PcG) family of proteins and plays an important role in the regulation of proliferation, senescence, cell cycle and apoptosis, chromosome stability, activation of gene transcription. METHODS:To clarify the roles of Bmi-1 in tumourigenesis and progression of gastric carcinomas, it was examined by immunohistochemistry (IHC) and real-time RT-PCR in gastric carcinomas, dysplasia, intestinal metaplasia (IM), and gastritis with a comparison of its expression with clinicopathological parameters of carcinomas. RESULTS:There was gradually increased Bmi-1 protein expression from gastritis, IM, dyplasia to carcinoma (p<0.001). Bmi-1 expression was positively linked to tumor size, depth of invasion, lymph node metastasis and worse prognosis of carcinomas (p<0.001), but not to age or sex of carcinoma patients (p>0.05). There was higher Bmi-1 protein expression in intestinal-type carcinomas than diffuse-type ones (p<0.001). At mRNA level, Bmi-1 protein expression was increased from gastritis, IM, dysplasia and carcinoma (p<0.001). Bmi-1 overexpression was observed in gastric carcinoma with larger diameter, deeper invasion, lymph node metastasis, and intestinal-type carcinoma (p<0.05). CONCLUSION:These findings indicate that up-regulated Bmi-1 expression is positively linked to pathogenesis, growth, invasion, metastasis and differentiation of gastric carcinomas. It was considered as a promising marker to indicate the aggressive behaviors and prognosis of gastric carcinomas.

OBJECTIVE:To detect the expressions of nuclear factor-Kappa B (NF-κB) and intercellular adhesion molecule-1 (ICAM-1) after hepatic ischemia-reperfusion injury so as to study the protective effect of Salvia in rats.METHODS:The hepatic ischemia-reperfusion rat model was established by the method of Nauta. The rats were randomly divided into 3 groups: sham operation (S), ischemia-reperfusion (I/R) and Salvia preconditioning (P). At 2, 8, 24 h after ischemia-reperfusion, immunohistochemistry was employed to detect the hepatic expressions of NF-κB and ICAM-1. Measurement of hepatic tissue myeloperoxidase (MPO) concentration was used to reflect the neutrophilic infiltration of hepatic tissue. And the injury severity of hepatic tissue was evaluated by the levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and the examinations of light microscopy and electron microscopy.RESULTS:The NF-κB level at 2 h (0.418 ± 0.039) post-reperfusion was significantly higher in the I/R group. And it decreased gradually at 8 h (0.308 ± 0.028) and 24 h (0.240 ± 0.032). And the ICAM-1 level increased at 2 h (0.367 ± 0.034) post-reperfusion, peaked at 8 h(0.451 ± 0.031)and began to decrease at 24 h (0.293 ± 0.025). The changes of MPO, ALT and AST were similar to ICAM-1. The above-mentioned indicators at different time-points were lower in the P group than those of the I/R group (P < 0.05), but higher than the S group (P < 0.05). The changes in ALT, MPO, NF-κB and ICAM-1 had a positive correlation (P < 0.01). Light and electron microscopy observations showed significant differences in different groups.CONCLUSION:The rat hepatic ischemia-reperfusion increases the expressions of NF-κB and ICAM-1 and they are involved in hepatic ischemia-reperfusion injury. Salvia significantly reduces the expressions of NF-κB and ICAM-1 in ischemia-reperfusion hepatic tissue and it may effectively protect the injury of hepatic ischemia-reperfusion.

Objective To construct human defensin-5(HD-5) gene into eukaryotic expression vector HD5-pPICZαA. Methods HD-5 was amplified from human cDNA by PCR. HD-5 gene product fragments and Pichia pastoris expression vector pPICZαA were double-digested by EcoRⅠand XbaⅠand connected by T4 DNA ligase. Ligation mixtures were transformed into competent E.coli Top10, and positive clones were screened with Zeccin. Transformants were identified by enzyme digesting, PCR and sequencing analysis. Results It was proved that HD-5 gene segment was inserted into pPICZαA correctly after identification with PCR enzyme cutting and sequencing analysis. Conclusion Eukaryotic expression vector HD5-pPICZαA was successfully constructed, which would provide a exprimental basis for further studyon the HD-5 expression in Pichia pastoris.

[Objective] To explore the relationship between the expression of CXCR4 and its ligand SDF-1 in gastric cancer and several clinicopathological factors. [Methods] The specimens were surgically resected from 62 patients with gastric cancer who underwent gastrectomy in The First Affiliated Hospital of Liaoning Medical College. CXCR4 and SDF-1 expression levels in gastric cancer specimens obtained from 62 patients were examined by immunohistochemistry. [Results] The staining intensity of CXCR4 and SDF-1 in cancer cells was significantly higher in intestinal-type than in diffuse-type gastric cancer. There was a significant correlation between the expression of CXCR4 and SDF-1 and liver metastasis and lymphatic metastasis in the intestinal-type cancer; however, this correlation was not found in the diffuse-type cancer. Furthermore, intestinal-type cancer cells that permeated the vascular or lymphatic channels as well as liver and lymph node metastases showed strong CXCR4 and SDF-1 staining. [Conclusion] The over-expressed CXCR4 and SDF-1 in intestinal-type cancer cells may be therapeutic targets for preventing lymphatic and hematogenous metastasis.