Serrated polyposis syndrome (SPS) is closely associated with the initiation and development of colorectal cancer (CRC), however, there is few research on SPS in China. Serrated polyps can be divided into hyperplastic polyps, sessile serrated polyps and traditional serrated polyps. The diagnosis standard of SPS is as following: (1) There are at least 5 serrated lesions in proximal colon, and diameter of more than 2 lesions is >10 mm; (2) The patient has one serrated polyp with family history of SPS; (3) More than 20 serrated polyps can be found in the entire large bowel. The risk of SPS is relatively high in the development of colorectal cancer and 25%-70% of the SPS patients is diagnosed with synchronous or metachronous colorectal cancer during following-up. The clinical characteristics of SPS include that patients are relatively old; no significant racial difference exists in the morbidity; patients have family history of colorectal cancer. The mutation of BRAF or KRAS gene, which induces colorectal cancer through the RAS-RAF-MAPK signaling pathway, is often found in SPS as well as CpG island methylation phenotype (CIMP) and microsatellite instability (MSI). The difference between SPS and traditional familial adenomatous polyposis (FAP) should be noted because of the different pathology mechanism, clinical characteristics and the risk of malignancy. Nowadays, the common technologies of detecting serrated polyps are auto-fluorescence imaging (AFI) and narrow-band imaging (NBI), whose detective rate is around 55%. The SPS patients are advised to undergo the resection of all the serrated polyps with diameter larger than 3-5 mm and receive the colonoscopy examination every 1 or 2 year. Not only the research about SPS is on the initiation step and the molecular mechanism is still unknown, but also the scholars do not come to achieve agreement about the risk of SPS in the malignancy of colorectal cancer, which is essential for further research therefore.
Adenoma ; genetics ; pathology ; therapy ; Adenomatous Polyposis Coli ; genetics ; pathology ; therapy ; Colonic Polyps ; Colonoscopy ; Colorectal Neoplasms ; genetics ; pathology ; therapy ; DNA Methylation ; Genes, ras ; Humans ; Microsatellite Instability ; Mutation ; Phenotype ; Proto-Oncogene Proteins B-raf ; Syndrome

OBJECTIVETo explore the effect of PI3K p85alpha gene silencing on the 5-fluorouracil (5-FU)-induced apoptosis of colorectal cancer cells.

METHODSThe PI3K p85alpha/RNAi transfected cells (PI3K p85alpha/RNAi-LoVo) were cultured in RPMI 1640 supplemented with 10% fetal calf serum and 500 microg/ml G418. The 50% inhibitory concentration (IC50) values of 5-FU (0.000625, 0.00125, 0.005, 0.01, 0.02, 0.04, 0.08, 0.16, 0.32 micromol/ml) were evaluated by MTT assay. Mitochondrial membrane potential was detected by JC-1 fluorescence, and Western blotting was used to analyze the expression of apoptotic proteins Bcl-6 and Bim.

RESULTSCompared with the untransfected LoVo cells, PI3K p85alpha/RNAi-LoVo showed obviously decreased IC(50) of 5-FU (P=0.000). The mitochondrial membrane potential of PI3K p85alpha/RNAi-LoVo cells was significantly lower than that of LoVo cells, suggesting that silencing PI3K p85alpha expression increased the sensitivity of LoVo cells to 5-FU. The expression of apoptotic protein Bcl-6 and Bim were significantly higher in PI3K p85alpha/RNAi-LoVo cells treated with 5-FU than LoVo cells (P=0.000).

CONCLUSIONPI3Kp85alpha gene silencing can significantly promote 5-FU-induced apoptosis of colorectal LoVo cells.

Apoptosis ; genetics ; Cell Line, Tumor ; Class Ia Phosphatidylinositol 3-Kinase ; genetics ; metabolism ; Colorectal Neoplasms ; genetics ; pathology ; Fluorouracil ; pharmacology ; Genetic Therapy ; methods ; Humans ; RNA Interference

OBJECTIVETo investigate the biological behavioral effects of specific siRNA expression plasmids targeted against c-Myc and vascular endothelial growth factor (VEGF) on human colorectal cancer cell line Volo.

METHODSThe expression plasmids with small interfering RNA (siRNA) aiming at c-Myc and VEGF were designed and constructed respectively, then transfected into Volo cells by eukaryocyte transfection technique. The protein expressions of c-Myc and VEGF were detected by Western blotting. Cellular proliferation, apoptosis, cycle distribution and invasion character were analyzed by tetrazolium bromide colorimetry (MTT), flow cytometry (FCM), TUNEL assay and matrigel invasion assay respectively.

RESULTSEnzymatic digestion and DNA sequencing confirmed that the c-Myc and VEGF specific siRNA expression plasmids were constructed successfully. After plasmids were transfected into cells, the protein expressions of c-Myc and VEGF were significantly down-regulated respectively as compared with control group (P<0.01). The cellular proliferation inhibitory rates in c-Myc siRNA group, VEGF siRNA group and c-Myc+VEGF group were (59.20+/-5.05)%, (32.31+/-3.48)% and (75.81+/-7.89)% respectively, which were higher than that in control group [(6.80+/-1.45)%] (all P<0.05). The cell apoptosis rate in above 3 groups were (40.50+/-4.37)%, (21.30+/-2.98)% and (62.59+/-9.66)% respectively, which were higher than that in control group [(2.90+/-0.36)%] (all P<0.05). The cell invasion rates in VEGF siRNA group and c-Myc+VEGF siRNA group were (7.34+/-3.65)% and (2.80+/-1.02)%, which were lower than that in control group [(18.57+/-7.46)%] (P<0.05). The effect of c-Myc+VEGF siRNA group was greater.

CONCLUSIONSThe specific siRNA efficiently silences the expression of c-Myc and VEGF, subsequently, suppresses the cell proliferation, triggers the cell apoptosis and inhibits the cell invasiveness in these transfected colorectal cancer Volo cells. In addition, the synergism of siRNA-c-Myc and siRNA-VEGF in transfected cells can be found.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; genetics ; pathology ; Genes, myc ; Genetic Therapy ; Humans ; RNA, Small Interfering ; Vascular Endothelial Growth Factor A ; genetics

The aim of the present study was to accurately evaluate the association of Sox2 expression with the survival of patients with digestive tract cancers. Relevant literatures were identified by comprehensively searching databases including the Pubmed, Embase, CBMdisc, and Wanfang (up to October 2014). A meta-analysis was performed to clarify the association between Sox2 expression and overall survival or clinicopathological parameters of patients with digestive tract cancers (esophageal, gastric, and colorectal cancers). The results showed a significant association between high Sox2 expression and poor overall survival in patients with digestive tract carcinomas (HR=1.55, 95% CI=1.04-2.31), especially for patients with esophageal cancer (HR=2.04, 95%CI=1.30-3.22), colorectal cancer (HR=1.40, 95% CI=1.04-1.89), and digestive tract adenocarcinoma (HR=1.80, 95% CI=1.12-2.89), for Europeans (HR=1.98, 95% CI=1.44-2.71) or patients who did not receive neoadjuvant treatment (HR=1.73, 95% CI=1.10-2.72). Furthermore, Sox2 over-expression was highly correlated with vascular invasion (OR=1.86, 95% CI=1.25-2.77) and poor differentiation (OR=1.88, 95% CI=1.14-3.08), especially in esophageal and colorectal cancers. In conclusion, Sox2 expression may serve as a novel prognostic factor for patients with digestive tract cancers. Over-expression of Sox2 that is correlated with vascular invasion and poor differentiation suggests poor outcomes of patients with digestive tract cancers.
Antineoplastic Agents ; therapeutic use ; Biomarkers, Tumor ; genetics ; metabolism ; Colorectal Neoplasms ; diagnosis ; drug therapy ; mortality ; pathology ; Esophageal Neoplasms ; diagnosis ; drug therapy ; mortality ; pathology ; Gastrointestinal Tract ; metabolism ; pathology ; Gene Expression ; Humans ; Neoadjuvant Therapy ; methods ; Neoplasm Grading ; Neoplasms, Vascular Tissue ; diagnosis ; drug therapy ; mortality ; secondary ; Prognosis ; SOXB1 Transcription Factors ; genetics ; metabolism ; Stomach Neoplasms ; diagnosis ; drug therapy ; mortality ; pathology ; Survival Analysis

Antibodies, Monoclonal ; therapeutic use ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; therapeutic use ; Cadherins ; genetics ; metabolism ; Carcinoma, Papillary ; pathology ; Cetuximab ; Colorectal Neoplasms ; drug therapy ; genetics ; pathology ; Consensus ; Dissent and Disputes ; Drug Delivery Systems ; Epithelial-Mesenchymal Transition ; Gastrointestinal Neoplasms ; classification ; pathology ; Genes, ras ; Humans ; Mutation ; Neoplasm Invasiveness ; Neoplastic Syndromes, Hereditary ; genetics ; metabolism ; pathology ; Neuroendocrine Tumors ; classification ; pathology ; Precancerous Conditions ; pathology ; Stomach Neoplasms ; genetics ; metabolism ; pathology

Antineoplastic Agents ; therapeutic use ; Colorectal Neoplasms ; classification ; drug therapy ; genetics ; pathology ; Humans ; Microsatellite Instability ; Mutation ; Polymorphism, Single Nucleotide ; Precision Medicine ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins B-raf ; genetics ; Proto-Oncogene Proteins p21(ras) ; ras Proteins ; genetics

This experimental study sought to find out the inhibitory effects of Ad-GFP-nm23-H1 on proliferation and metastasis of human colorectal carcinoma cell line Lovo, and, further, to gain an insight into some theoretical and methodical basis for instituting nm23-H1 gene therapy of cancers. MTT assay and Transwell chamber were used to detect the rates of proliferation and invasion as well as the adhesion of Lovo cells in vitro. The results demonstrated that the proliferation inhibition rates of Lovo cells treated with Ad-GFP-nm23-H1 of 10(10) PFU/ml, 10(9) PFU/ml and 10(8) PFU/ml were 84.9% +/- 1.51%, 48.5% +/- 7.23% and 22.5% +/- 5.47%, that the adherence inhibition rates of Lovo cells treated with Ad-GFP-nm23-H1 of 10(10) PFU/ml, 10(9) PFU/ml and 10(8) PFU/ml were 70.3% +/- 2.40%, 60.1% +/- 5.68% and 18.5% +/- 3.61%, and that the invasiveness inhibition rates of Lovo cells treated with Ad-GFP-nm23-H1 of 10(10) PFU/ml, 10(9) PFU/ml and 10(8) PFU/ml were 83.2% +/- 5.71%, 52.2% +/- 6.94% and 28.1% +/- 8.21%. These data suggested that Ad-GFP-nm23-H1 exerted significant inhibitory effects on the proliferation and metastasis of human colorectal carcinoma cell line Lovo in a dose-dependent way.
Adenoviridae ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; genetics ; pathology ; Gene Expression Regulation, Neoplastic ; Genetic Therapy ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; NM23 Nucleoside Diphosphate Kinases ; genetics ; metabolism

OBJECTIVETo study FasL gene expression in colorectal carcinoma and its influences on biological behaviour of colorectal cancer and hepatic metastasis.

METHODSFasL gene expressions were examined with RT-PCR technique in the primary locus of colorectal cancer, mucosa adjacent to cancer, and hepatic metastasis. HR-8348 cells of human rectal cancer cell line were transfected with FasL cDNA. Cell growth suppression rates and cell response to 5-FU and carboplatin were observed and analysed with MTT method.

RESULTSFasL gene expressions were detected in the primary site of colorectal cancer (n = 58), cancer adjacent mucosa (n = 58), and hepatic metastasis (n = 28). The positive rate of FasL expression was 24% (14/58), 14% (8/58), 100% (28/28), respectively, in primary site, tumor adjacent mucoca and hepatic metastasis. FasL expression rate in hepatic metastasis was higher than that in the primary site (chi2 = 43.49, P < 0.01) and tumor adjacent mucosa (chi2 = 57.66, P < 0.01). In a group of patients with hepatic metastasis, FasL expression rate in primary site was higher than that in patients without hepatic metastasis (chi2 = 3.96, P < 0.05). In vitro experiment, positive expression of FasL was found in transfected HR-8348 cells. When 5-FU or carboplatin was added, there was a significant difference in growth suppression rate between FasL positive and control cancer cells (t = 9.02, t = 11.93, P < 0.01). Under same concentration of chemotheraputic agent, survival rate of FasL positive HR-8348 cells was higher than that of control cells.

CONCLUSIONSFasL positive cancer cells have more powerful resistance to chemotheraputic drugs. Expression of FasL gene in colorectal cancer cells is related with immune evasion to escape killing by immune cells, showing stronger drug-resistance, and it facilitates hepatic metastasis.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; drug therapy ; genetics ; pathology ; Drug Resistance, Neoplasm ; Fas Ligand Protein ; Female ; Humans ; Liver Neoplasms ; secondary ; Male ; Membrane Glycoproteins ; genetics ; Middle Aged ; Transfection

BACKGROUNDIn most colorectal carcinomas, the level of phospholipase C (PLC)-gamma 1 expression is greatly elevated. Increased expression of PLC-gamma 1 may play an important role in colon carcinogenesis, but the mechanism is not well known. The aim of this study was to evaluate the role of PLC-gamma 1 in colon carcinogenesis by using recombinant lentivirus that stably suppressed the PLC-gamma 1 expression in human colorectal carcinoma LoVo cells.

METHODSRecombinant lentivirus producing PLC-gamma 1 siRNA were prepared. After LoVo cells were transduced by each lentivirus, stably transduced cells were selected by Blasticidin. The protein and mRNA expression of PLC-gamma 1 were examined by Western-blot and reverse transcription-polymerase chain reaction (RT-PCR) analysis, and the effects of the lentivirus on the cell adhesion, migration and apoptosis were analyzed.

RESULTSStable LoVo cell line deficient in PLC-gamma 1, was established. Notably, PLC-gamma 1 was silenced without affecting the levels of other subtypes of PLC so that the role of PLC-gamma 1 in colon carcinogenesis could be examined. Silencing of endogenous PLC-gamma 1 resulted in efficient inhibition of the adhesion and migration of LoVo cells in vitro and a great increase of 5-fluorouracil induced apoptosis (30%-40%) of LoVo cells.

CONCLUSIONSPLC-gamma 1 may play an important role in metastasis and anti-apoptosis in human colorectal carcinomas.

Apoptosis ; drug effects ; Cell Adhesion ; Cell Line, Tumor ; Colorectal Neoplasms ; pathology ; therapy ; Fluorouracil ; pharmacology ; Humans ; Laminin ; antagonists & inhibitors ; genetics ; Lentivirus ; genetics ; Phospholipase C gamma ; antagonists & inhibitors ; genetics ; physiology ; RNA, Small Interfering ; therapeutic use

The p53 gene has a significant role in controlling genomic stability of cancer. The purpose of this study was to evaluate the tumor response of allograft colorectal tumor treated with Ad5CMV-p53 in a syngeneic rat model. Two weeks after the inoculation of WB-2054-M5 tumor cells in the flank of rats, rats were randomly assigned by tumor size to one of three groups (n=18 in each): phosphate buffered saline (PBS), Ad5CMV, and Ad5CMV-p53. Recombinant adenovirus or PBS was administered through intratumoral injection at three divided doses every other day for 4 weeks. Apoptosis of the tumors was evaluated using TUNEL assay. After 2 and 4 weeks of treatment, the volume (cm3) of tumors in PBS, Ad5CMV, and Ad5CMV-p53 was as follows: 2 week: 1.66 +/-0.43, 1.40 +/-0.47, 0.75 +/-0.26 (p<0.001), 4 week: 4.41 +/-0.88, 3.93 +/-1.86, 2.33 +/-0.51 (p<0.001). Tumor growth showed no statistically significant difference between the PBS and Ad5CMV groups (6-week vol. p=0.32). The TUNEL assay results revealed more apparent apoptotic cells in Ad5CMV-p53-treated tumors than in other groups. Growth of allograft colorectal cancer in the syngeneic rat model was significantly suppressed by intratumoral Ad5CMV-p53 gene therapy. These results demonstrate that gene replacement therapy with p53 may provide a novel modality of treatment in conjunction with other present treatments for metastatic colorectal cancer.
Adenocarcinoma/genetics/pathology/therapy ; Adenoviridae/*genetics ; Animals ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival/genetics ; Colorectal Neoplasms/*genetics/pathology/*therapy ; Disease Models, Animal ; Female ; Gene Therapy/*methods ; Gene Transfer Techniques ; Men ; Protein p53/*genetics/*therapeutic use ; Rats ; Rats, Inbred WF ; Recombinant Proteins/therapeutic use ; Research Support, Non-U.S. Gov't ; Transplantation, Isogeneic ; Treatment Outcome