Medical Library

Over-expressed and truncated midkines promote proliferation of BGC823 cells in vitro and tumor growth in vivo.

AIM: To determine whether midkine (MK) and its truncated form (tMK) contribute to gastric tumorigenesis using in vitro and in vivo models. METHODS: Human MK and tMK plasmids were constructed and expressed in BGC823 (a gastric adenocarcinoma cell line) to investigate the effect of over-expressed MK or tMK on cell growth and turmorigenesis in nude mice. RESULTS: The growth of MK-transfected or tMK-transfected cells was significantly increased compared with that of the control cells, and tMK-transfected cells grew more rapidly than MK-transfected cells. The number of colony formation of the cells transfected with MK or tMK gene was larger than the control cells. In nude mice injected with MK-transfected or tMK-transfected cells, visible tumor was observed earlier and the tumor tissues were larger in size and weight than in control animals that were injected with cells without the transfection of either genes. CONCLUSION: Over-expressed MK or tMK can promote human gastric cancer cell growth in vitro and in vivo, and tMK has greater effect than MK. tMK may be a more promising gene therapeutic target compared with MK for treatment of malignant tumors.

Wang QL, Wang H, Zhao SL, Huang YH, Hou YY.

Immunology and Reproductive Biology Lab, Medical School and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, Jiangsu Province, China. yayihou@nju.edu.cn.

Effect of JIANPI HUOXUE decoction on inflammatory cytokine secretion pathway in rat liver with lipopolysaccharide challenge.

AIM: To evaluate the effect of Chinese traditional medicinal prescription, JIANPI HUOXUE decoction (JHD) on cytokine secretion pathway in rat liver induced by lipopolysaccharide (LPS). METHODS: Twenty-four male SD rats were divided into normal group (n = 4), model group (n = 10) and JHD group (n = 10) randomly. Rats in model group and JHD group were administrated with normal saline or JHD via gastrogavage respectively twice a day for 3 d. One hour after the last administration, rats were injected with LPS via tail vein, 50 mug/kg. Simultaneously, rats in normal group were injected with equivalent normal saline. After LPS stimulation for 1.5 h, serum and liver tissue were collected. Pathological change of liver tissues was observed through hematoxylin-eosin (H.E.) staining. Tumor necrosis factor alpha (TNF-alpha) in serum were assayed by enzyme linked immunosorbent assay (ELISA). The protein expression of TNF-alpha, phosphorylated inhibit-kappaB (p-IkappaB) and CD68 in liver were assayed by Western blot. The distribution of CD68 protein in liver was observed through immunohistochemical staining. The mRNA expression of TNF-alpha, interleukin-6 (IL-6), CD14, toll-like receptor 2 (TLR2) and TLR4 in liver were assayed by real-time RT-PCR. RESULTS: Predominant microvesicular change, hepatocyte tumefaction and cytoplasm dilution were observed in liver tissues after LPS administration as well as obvious CD68 positive staining in hepatic sinusoidal. After LPS stimulation, serum TNF-alpha (31.35 +/- 6.06 vs 12225.40 +/- 9007.03, P < 0.05), protein expression of CD68 (1.13 +/- 0.49 vs 3.36 +/- 1.69, P < 0.05), p-IkappaB (0.01 +/- 0.01 vs 2.07 +/- 0.83, P < 0.01) and TNF-alpha (0.27 +/- 0.13 vs 1.29 +/- 0.37, P < 0.01) in liver and mRNA expression of TNF-alpha (1.96 +/- 2.23 vs 21.45 +/- 6.00, P < 0.01), IL-6 (4.80 +/- 6.42 vs 193.50 +/- 36.36, P < 0.01) and TLR2 (1.44 +/- 0.62 vs 4.16 +/- 0.08, P < 0.01) in liver were also increased significantly. These pathological changes were all improved in JHD group. On the other hand, TLR4 mRNA (1.22 +/- 0.30 vs 0.50 +/- 0.15, P < 0.05) was down-regulated and CD14 mRNA increased but not significantly after LPS stimulation. CONCLUSION: JHD can inhibit cytokine secretion pathway induced by LPS in rat liver, which is probably associated with its regulation on CD68, p-IkappaB and endotoxin receptor TLR2.

Peng JH, Hu YY, Cheng Y, Han C, Xu LL, Feng Q, Chen SD, Tao Q, Li HS, Li XM.

Institute of Liver Disease, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, No. 528, Zhangheng Road, Shanghai 201203, China. yyhu@citiz.net.

Immune-mediated anti-neoplastic effect of intratumoral RSV envelope glycoprotein expression is related to apoptotic death of tumor cells but not to the size of syncytia.

AIM: To promote the development of improved tumor vaccination strategies relying on the intratumoral expression of viral fusogenic membrane proteins, we elucidated whether the size of syncytia or the way tumor cells die has an effect on the therapeutic outcome. METHODS: In two syngeneic subcutaneous murine colon cancer models we assessed the anti-neoplastic effect on vector-treated and contralateral untreated tumors. RESULTS: Intratumoral injection of a replication-defective adenovirus encoding respiratory syncytial virus fusion protein (RSV-F) alone (Ad.RSV-F) or together with the attachment glycoprotein RSV-G (Ad.RSV-F/G) led to a significant growth reduction of the vector-treated and contralateral untreated tumors. The treatment response was associated with a strong tumor-specific CTL response and significantly improved survival with medians of 46 d and 44 d, respectively. Intratumoral injection of Ad.RSV-G or a soluble RSV-F encoding adenovirus (Ad.RSV-F(sol)) had no significant anti-neoplastic effect. The median survival of these treatment groups and of Ad.Null-treated control animals was about 30 d. CONCLUSION: Although in vitro transduction of colon cancer cell lines with Ad.RSV-F/G resulted in about 8-fold larger syncytia than with Ad.RSV-F, the in vivo outcome was not significantly different. Transduction of murine colon cancer cell lines with Ad.RSV-F or Ad.RSV-F/G caused apoptotic cell death, in contrast to transduction with Ad.RSV-G or Ad.RSV-F(sol), suggesting an importance of the mode of cell death. Overall, these findings provide insight into improved tumor vaccination strategies relying on the intratumoral expression of viral fusogenic membrane proteins.

Hoffmann D, Grunwald T, Bayer W, Wildner O.

Ruhr-University Bochum, Institute of Microbiology and Hygiene, Department of Molecular and Medical Virology, Bldg. MA, Rm. 6/40, Bochum 44801, Germany. oliver.wildner@ruhr-uni-bochum.de.

Effects of two novel nucleoside analogues on different hepatitis B virus promoters.

AIM: To explore the effects of the nucleoside analogues beta-L-D4A and beta-LPA on hepatitis B virus (HBV) promoters. METHODS: Four HBV promoters were amplified by polymerase chain reaction (PCR) and subcloned into the expression vector pEGFP-1. The four recombinants controlled by HBV promoters were confirmed by restriction analysis and sequencing. Human hepatoma HepG2 cells transfected with the recombinant plasmids were treated with various concentrations of beta-L-D4A and beta-LPA. Then, enhanced green fluorescent protein (EGFP)-positive cells were detected by fluorescence microscopy and using a fluorescence activated cell sorter (FACS). RESULTS: Four HBV promoters were separately obtained and successfully cloned into pEGFP-1. Expression of EGFP under the control of the surface promoter (Sp) and the X promoter (Xp) was inhibited by beta-L-D4A in a dose-dependent manner, while expression of EGFP under the control of the core promoter (Cp) and Xp was inhibited by beta-LPA in a dose-dependent manner. CONCLUSION: The two novel nucleoside analogues investigated here can inhibit the activities of HBV promoters in a dose-dependent manner. These findings may explain the mechanisms of action by which these two novel compounds inhibit HBV DNA replication.

He XX, Lin JS, Chang Y, Zhang YH, Li Y, Wang XY, Xu D, Cheng XM.

Institute of Liver Diseases, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China. jslin@tjh.tjmu.edu.cn.

Comparative genomic hybridization analysis of genetic aberrations associated with development of esophageal squamous cell carcinoma in Henan, China.

AIM: To characterize cytogenetic alterations in esophageal squamous cell carcinoma (ESCC) and its metastasis. METHODS: A total of 37 cases of primary ESCC and 15 pairs of primary ESCC tumors and their matched metastatic lymph nodes cases were enrolled from Linzhou, the high incidence area for ESCC in Henan, northern China. The comparative genomic hybridization (CGH) was applied to determine the chromosomal aberrations on the DNA extracted from the frozen ESCC and metastatic lymph node samples from these patients. RESULTS: CGH showed chromosomal aberrations in all the cases. In 37 cases of primary ESCC, chromosomal profile of DNA copy number was characterized by frequently detected gains at 8q (29/37, 78%), 3q (24/37, 65%), 5p (19/37, 51%); and frequently detected losses at 3p (21/37, 57%), 8p and 9q (14/37, 38%). In 15 pairs of primary ESCC tumors and their matched metastatic lymph node cases, the majority of the chromosomal aberrations in both primary tumor and metastatic lymph node lesions were consistent with the primary ESCC cases, but new candidate regions of interest were also detected. The most significant finding is the gains of chromosome 6p with a minimum high-level amplification region at 6p12-6q12 in 7 metastatic lymph nodes but only in 2 corresponding primary tumors (P = 0.05) and 20p with a minimum high-level amplification region at 20p12 in 11 metastatic lymph nodes but only in 5 corresponding primary tumors (P < 0.05). Another interesting finding is the loss of chromosome 10p and 10q in 8 and 7 metastatic lymph nodes but only in 2 corresponding primary tumors (P < 0.05). CONCLUSION: Using the CGH technique to detect chromosomal aberrations in both the primary tumor and its metastatic lymph nodes of ESCC, gains of 8q, 3q and 5p and loss of 3p, 8p, 9q and 13q were specifically implicated in ESCC in Linzhou population. Gains of 6p and 20p and loss of 10pq may contribute to the lymph node metastasis of ESCC. These findings suggest that the gains and losses of chromosomal regions may contain ESCC-related oncogenes and tumor suppressor genes and provide important theoretic information for identifying and cloning novel ESCC-related oncogenes and tumor suppressor genes.

Qin YR, Wang LD, Fan ZM, Kwong D, Guan XY.

Department of Clinical Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China. yanruqin@163.com.

Crosstalk between tumor cells and microenvironment via Wnt pathway in colorectal cancer dissemination.

Invasion and metastasis are the deadly face of malignant tumors. Considering the high rate of incidence and mortality of colorectal cancer, it is critical to determine the mechanisms of its dissemination. In the parallel investigation of the invasive front and tumor center area of colorectal cancer (CRC), observation of heterogeneous beta-catenin distribution and epithelial-mesenchymal transition (EMT) at the invasive front suggested that there might be a crosstalk between tumor cells and the tumor microenvironment. Wnt signaling pathway is also involved in the cancer progression due to its key role in CRC tumorigenesis. Moreover, in recent years, there is increasing evidence that the regulators of microenvironment, including extracellular matrix, growth factors and inflammatory factors, are associated with the activation of Wnt pathway and the mobility of tumor cells. In this review, we will try to explain how these molecules trigger metastasis via the Wnt pathway.

Huang D, Du X.

270 Dong An Road, Department of Pathology, Cancer Hospital, Fudan University, Shanghai 200032, China. dx2008cn@yahoo.com.cn.

Role of chemotherapy and novel biological agents in the treatment of elderly patients with colorectal cancer.

Patients older than 65 years are the fastest growing segment of the cancer population. It is estimated that within 20 years over 75% of cases and 85% of deaths from colorectal cancer (CRC) will be in this setting. Concerns about cancer treatment in the elderly relate to comorbidities, which increase proportionally with age, physiological changes associated with aging which may influence drug metabolism and toxicity, and diminishing life expectancy, which particularly impacts decisions surrounding the benefits of adjuvant therapies. Over the last 10 years, significant improvements in the treatment of advanced CRC with combination therapy have been made. The randomized trials which have defined these improvements did not exclude elderly patients. However, the median age of patients in these trials has generally been approximately 60 years. Thus, it appears that some degree of selection is involved with younger and presumably fitter patients being the subjects in most of the pivotal trials. The availability of new molecularly targeted agents and newly improved existing agents has expanded the range of treatment options available. This variety gives greater flexibility in dealing with different subsets of patients, such as the elderly. However, some fit elderly patients seem to tolerate combination therapy reasonably well, while studies on unfit elderly subjects are needed.

Rosati G, Bilancia D.

Medical Oncology Unit, S. Carlo Hospital, Via P. Petrone 1, Potenza 85100, Italy. rosatiger@yahoo.com.

Colorectal cancer risk in Crohn\’s disease.

There is recognized increased risk for colorectal cancer in patients with inflammatory bowel disease, particularly in long-standing and extensive ulcerative colitis. There also appears to be an increased rate of intestinal cancer in Crohn\’s disease, including both colon and small bowel sites. In Crohn\’s disease, evidence suggests that detection of colorectal cancer may be delayed with a worse prognosis. Some risk factors for cancer in Crohn\’s disease include the extent of inflammatory change within the colon and the presence of bypassed or excluded segments, including rectal \”stump\” cancer. In addition, the risk for other types of intestinal neoplasms may be increased in Crohn\’s disease, including lymphoma and carcinoid tumors. Earlier detection of colorectal cancer based on colonoscopy screening and surveillance may be achieved but, to date, this has not translated into a positive survival benefit. Moreover, newer staining methods and evolving micro-endoscopic techniques show promise, but have not significantly altered management. Future research should focus on development of molecular or other bio-markers that might predict future dysplasia or cancer development in Crohn\’s disease.

Freeman HJ.

Department of Medicine (Gastroenterology), University of British Columbia, UBC Hospital, 2211 Wesbrook Mall, Vancouver, BC, Canada V6T 1W5, Canada. hugfree@shaw.ca.

Intragastric injection of botulinum toxin for the treatment of obesity. Where are we?

Obesity has reached epidemic proportions particularly in western countries. Most non-surgical treatments of this condition are disappointing. Since 2005, several studies evaluating the effect of Botulinum Toxin type A (BT-A) in gastric antrum by means of endoscopy for the treatment of obesity have been published. This treatment modality was based on the observation that gastric injection of BT-A in laparatomized rats induced a significant reduction of food intake and body weight. Nowadays, 6 studies have been published yielding conflicting results. Differences in selection of patients, doses of BT-A, method of administration of the toxin and instruments of evaluation of some parameters among these studies may be the cause of divergent results. We discuss herein some important features of these studies pointing out on differences among them. At the same time, based on the knowledge of physiological characteristics of normal and abnormal gastric function related with feeding, we discuss the probable causes of failure observed in these trials. Finally, we give some guidelines concerning the way that future research in this field may follow, not without calling attention to disadvantages of this treatment.

Garcia-Compean D, Maldonado Garza H.

Department of Gastroenterology, Faculty of Medicine, University Hospital, Ave Madero y Gonzalitos, Col Mitras Centro, Monterrey 64700, Mexico. digarciacompean@prodigy.net.mx.

Hemobilia due to hepatic artery aneurysm as the presenting sign of fibro-muscular dysplasia.

Fibro-muscular dysplasia (FMD) is a rare but well documented disease with multiple arterial aneurysms. The patients, usually women, present with various clinical manifestations according to the specific arteries that are affected. Typical findings are aneurysmatic dilatations of medium-sized arteries. The renal and the internal carotid arteries are most frequently affected, but other anatomical sites might be affected too. The typical angiographic picture is that of a \”string of beads\”. Common histological features are additionally described. Here we present a case of a 47-year-old woman, who was hospitalized due to intractable abdominal pain. A routine work-up revealed a liver mass near the portal vein. Before a definite diagnosis was reached, the patient developed massive upper gastrointestinal bleeding. In order to control the hemorrhage, celiac angiography was performed revealing features of FMD in several arteries, including large aneurysms of the hepatic artery. Active bleeding from one of these aneurysms into the biliary tree indicated selective embolization of the hepatic artery. The immediate results were satisfactory, and the 5 years follow-up revealed absence of any clinical symptoms.

Shussman N, Edden Y, Mintz Y, Verstandig A, Rivkind AI.

Department of General Surgery, Hadassah Hebrew University Medical Center, POB 12000, Jerusalem 91120, Israel. noams@hadassah.org.il.