Effect of feeding fermentable fiber on synthesis of total and mucosal protein in the intestine of the growing pig

In a previous study, a reduced efficiency of ileal digestible threonine (THR) use for body protein deposition was observed in growing pigs when pectin was included in the diet. This response was not due to increased physical endogenous ileal THR loss. Our aim was to explore the contribution of diet-induced increases in protein synthesis in the colon, especially mucins, to dietary THR requirements. Twelve barrows (21 kg mean BW) were fed either a cornstarch–soybean meal-based diet (Control) or Control with 12% pectin (Pectin). Pigs were given intravenously 1.5 mmol/kg BW of L-1-¹³C valine (40 mol%) to measure fractional and absolute synthesis rates (FSR, ASR, respectively) of mucosal and whole intestinal protein in the jejunum and colon. Dietary pectin inclusion increased plasma levels of glucose, isoleucine and glutamine (P < 0.05) but had no effect on insulin or urea nitrogen (P > 0.10). There were no differences in FSR and ASR of whole intestinal protein in jejunum and colon (P > 0.10). The FSR of mucosal proteins in colon, not in jejunum, was increased with dietary pectin supplementation (P < 0.05). Assuming mucosal protein mass is constant, these results imply that the higher protein synthesis in colon mucosa contributes to the reduced THR efficiency observed in pectin-supplemented diet.     

Up/down-regulation of miR-21 changes biological function of colon can-cer cells and sensitivity to cetuximab

AIM: To explore the effects of miR-21 on biological behavior of colon cancer cells and their sensitivity to epidermal growth factor receptor monoclonal antibody cetuximab. METHODS: Lentiviral vectors were constructed to generate up- and down-regulations of miR-21 lentiviruses (LV-miR-21 and LV-anti-miR-21, respectively), and the corresponding negative control viruses (LV-miR-21 NC and LV-anti-miR-21 NC, respectively) were also constructed. The viruses were used to infect human colon cancer RKO cells. The changes of the miR-21 expression level, the cell proliferation, the colony-forming ability, the cell apoptosis and the sensitivity of the cells to cetuximab were detected by real-time PCR, MTT assay, soft agar colony assay, flow cytometry and CCK-8 assay. RESULTS: The lentivirus titers of LV-miR-21, LV-miR-2 NC, LV-anti-miR-21 and LV-anti-miR-21 NC were 3.0×1012 TU/L, 6.0×1011 TU/L, 2.0×1012 TU/L and 8.0×1011 TU/L, respectively. The infection efficiency was over 80% by the observation of green fluorescence. The miR-21 expression level, the cell proliferation, and the colony-forming ability in LV-miR-21 group were significantly higher than those in LV-anti-miR-21 group. The early apoptotic rate and the inhibitory rate of cetuximab for the cells in LV-anti-miR-21 group were higher than those in LV-miR-21 group. CONCLUSION: miR-21 promotes the proliferation of colon cancer cells. Down-regulation of miR-21 enhances the sensitivity of the colon cancer cells to the targeted therapy drug cetuximab.     

地方猪种对纤维饲料利用的研究——Ⅱ.消化道形态与大肠内容物重量的变化

以三元杂种猪为对照,研究了东北民猪在3种NDF水平(0、10、20％)下消化道形态及后肠内容物鲜重的变化。试验从活重30千克开始,80千克结束时,每组在最后一次给食后的2、4、8、12或16小时随机屠宰2头。电击晕死后立即开膛取出内脏,分为胃、小肠、盲肠和结肠4段进行测量。结果表明猪对饲粮纤维含量的增加从胃肠道形态上发生明显的适应性变化,消化道长度、鲜重增加,尤其结肠长度增加明显,胃、小肠和结肠的鲜重显著地高于基础饲粮组。民猪胃重和结肠鲜重及长度的增加比对照杂种猪明显,消化道总鲜重及其占空体重的比例显著地高于对照杂种猪。随NDF水平的提高,大肠内容物鲜重明显增加,其中民猪更为突出,后肠是吸收水分的主要场所。     

Study on expression of long non-coding RNA in colon cancer tissues and adjacent tissues

AIM: To screen the differentially expressed long non-coding RNA (lncRNA) in colon cancer, and to explore its expression in colon cancer tissues and adjacent tissues. METHODS: The "Colon adenocarcinoma:Person neoplasm cancer status" which consisted of 36 cases of colon cancer tissues and 29 cases of normal colonic tissues was downloaded from the lncRNAtor database. The candidate genes were selected from these differentially expressed lncRNAs based on artificial criterion (P<0.01; fold change ≥ 2 or<0.5) and then validated by real-time PCR in 60 pairs of colon cancer tissues and adjacent tissues. RESULTS: A total of 50 lncRNAs were differentially expressed in colon cancer tissues, including 28 up-regulated and 22 down-regulated (P<0.01). The verifying results displayed that HNF1A-AS1 and ZDHHC8P1 were up-regulated (P<0.01), and SUZ12P expression was down-regulated (P<0.05), but the expression of AC069513.3 was not statistically significant between colon cancer tissues and adjacent tissues. The abilities of HNF1A-AS1, ZDHHC8P1, SUZ12P and AC069513.3 to discriminate the colon cancer from normal adjacent tissue by the ROC curve with an AUC of 0.729 (sensitivity 78%, specificity 67%), 0.617 (sensitivity 68%, specificity 55%), 0.689 (sensitivity 66%, specificity 55%) and 0.518 (sensitivity 52%, specificity 48%) were observed. CONCLUSION: Long non-coding RNA HNF1A-AS1 and ZDHHC8P1 are up-regulated and SUZ12P is down-regulated in colon cancer tissues, suggesting that they may be involved in the pathogenesis of colon cancer.     

Cellulose and calcium lower the incidence of chemically-induced colon tumors in rats

In a 30-week preliminary study and a follow-up 22-week study (2 × 2 factorial), dimethylhydrazine (DMH)injections effectively induced colon tumors in Fischer-344rats. How this incidence of colon tumors might be affected by cellulose (preliminary study) or by calcium and folic acid (follow-up study) was examined. Cellulose in the dietappeared to provide some protection against DMH-induced colon tumors, but the protective effect of calcium was moreevident; normal levels of calcium (500 mg per 100 g diet), but not of folic acid (0.1 mg per 100 g diet), provided protection against colon tumors. The effect due to calciumwas observed whether viewed in terms of total number of tumors (p lt 0.01) or number of tumors per tumor-bearing rat(p < 0.01).     

Effect of 7-hydroxyisoflavone on HCT116 cells proliferation by Id1

AIM:To investigated the effect of 7-hydroxyisoflavone (7-HIF) on the proliferation, apoptosis and stem-like cell feature of colorectal cancer cells. METHODS:The effect of 7-HIF on the proliferation of HCT116 cells was detected by WST-1 assay and colony formation assay. The effects of 7-HIF on the cell cycle distribution and apoptosis in the HCT116 cells were analyzed by flow cytometry. The expression of cell-cycle related proteins and the stemness related proteins was determined by Western blot. RESULTS:After treated with 7-HIF (200 μmol/L), the viability of HCT116 cells was inhibited, and the size and number of the colony were decreased as compared with control group (P<0.05). The G₀/G₁ phase of the cell cycle was increased. The proportion of S phase was decreased and the cells were mainly arrested in G₀/G₁ phase. The apoptotic rate of HCT116 cells was 21.4%, which was significantly higher than that in the control group (1.1%). The results of Western blot revealed that the expression of inhibitor of differentiation 1(Id1) was significantly decreased (P<0.05). The expression of cell cycle markers cyclin D1 and cyclin E, the proliferative markers survivin and PCNA, and stem cell markers CD133, ALCAM and EpCAM were all down-regulated by 7-HIF treatment (P<0.05). CONCLUSION:7-HIF inhibits the proliferation and induces the apoptosis of colorectal cancer cells, and inhibits the stem-like cell feature, which may be related to Id1 inhibition.     

High mobility group box 1 protein promotes proliferation,migration and invasion of colon cancer cells

AIM: To investigate the expression of high mobility group box 1 protein (HMGB1) in colon can-cer cells, and to determine its regulatory roles in colon cancer cell proliferation, migration and invasion. METHODS: qPCR and Western blot were used to quantify the mRNA and protein expression levels of HMGB1 in human colon cancer SW620 cells and normal colonic epithelial FHC cells. HMGB1 shRNA was transfected into the SW620 cells to establish the stable HMGB1-downregulating colon cancer cells (shHMGB1 group), and negative control (shNC) group and blank control (blank) group were also set up. The proliferation, migration and invasion of the cells were determined by CCK-8 assay, colony formation experiment and Transwell chamber assays. Western blot was used to determine the protein levels of p-ERK, ERK, c-Myc, matrix metalloproteinase (MMP)-2/9, E-cadherin, N-cadherin, Bcl-2 and Bax. RESULTS: Both of the mRNA and protein levels of HMGB1 in colon cancer cells were higher than those in the normal colonic epithelial cells (P<0.05). HMGB1 gene was successfully knocked down in SW620 cells. Compared with blank group and shNC group, the proliferation, migration and invasion abilities of the cells in shHMGB1 group were significantly inhibited (P<0.05). The protein levels of MMP-2, MMP-9, N-cadherin, c-Myc, Bcl-2 and p-ERK were reduced notably, while the expression of Bax protein was increased (P<0.05) in shHMGB1 group compared with shNC group and blank group.CONCLUSION: HMGB1 effectively promotes the proliferation, migration and invasion of colon cancer cells through ERK/c-Myc signaling pathway.