Expression of Wnt and integrin pathways in colorectal laterally spreading tumors and their correlation with endoscopic subtypes / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of Wnt and integrin pathways in colorectal laterally spreading tumors (LSTs) and their correlation with the different endoscopic subtypes of LSTs to better understand the special growth mechanism of LSTs.</p><p><b>METHODS</b>Fifty-two patients with colorectal LSTs were randomly selected from the cases diagnosed between January 1, 2010 and June 10, 2015 in our hospital, including 37 of nodular mixed type (LST-G-M), 60 of homogeneous type (LST-G-H), 5 of flat elevated type (LST-NG-FE), and 4 of pseudodepressed type (LST-NG-PD). The expression of β-catenin, phospho- GSK-3β, paxillin and ILK in 52 colorectal LSTs and 15 protruded adenomas (PAs) were investigated by immunohistochemical staining. The correlation of β-catenin, phospho-GSK-3β, paxillin and ILK expressions among the endoscopic subtypes of LSTs were analyzed.</p><p><b>RESULTS</b>β-catenin expression was significantly higher in LSTs than in Pas (P<0.05). β-catenin, phospho-GSK-3β, paxillin and ILK expressions were significantly higher in LST-NG-PD than in Pas (P<0.05). The expressions of β-catenin, phospho-GSK-3β and ILK expression were significantly correlated in LSTs (P<0.05) but not in PAs (P>0.05).</p><p><b>CONCLUSION</b>The macroscopic feature of LST-NG-PD may result from a special mechanism of development distinct from other endoscopic subtypes; ILK may play a role in regulating Wnt signaling in LSTs.</p>

5-aza-2'-deoxycytidine-induced inhibition of CDH13 expression and its inhibitory effect on methylation status in human colon cancer cells in vitro and on growth of xenograft in nude mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To determine the inhibitory effect of 5-aza-2'-deoxycytidine (5-Aza-CdR) on the growth of human colon carcinoma cells and xenografts in nude mice, to observe its effect on CDH13 gene expression and methylation in the xenografts, and to explore the possible mechanisms.</p><p><b>METHODS</b>Human colon carcinoma cell line HCT116 cells were treated with 5-Aza-CdR, and the cell morphology was observe by phase contrast microscopy. The cell growth was assessed by MTT assay. A tumor-bearing mouse model was generated by subcutaneous inoculation of human colon carcinoma HCT116 cells into nude mice. The tumor growth in the nude mice was observed, the CDH13 gene expression and its methylation status in the tumors were detected using methylation specific PCR (MSP), RT-PCR, Western blotting and immunohistochemistry.</p><p><b>RESULTS</b>After treatment with 5-Aza-CdR, the inhibition rate of the growth of cultured HCT116 cells was increased as the concentration was increasing. The growth of the xenografts in nude mice was significantly inhibited, and the methylated CDH13 gene was reactivated. After 4 weeks of 5-Aza-CdR treatment, no significant difference was found between the body weights of nude mice in the 5-Aza-CdR group [(18.06 ± 1.29) g] and control group [(17.07 ± 0.84) g], (P > 0.10), and the average volume of xenografts of the 5-Aza-CdR group was (907.00 ± 87.29) mm(3), significantly smaller than the (1370.93 ± 130.20) mm(3) in the control group (P < 0.005). No expression of CDH13 gene was found in the control group. The expression of CDH13 gene in the 5-Aza-CdR group was increased along with the increasing concentration of 5-Aza-CdR.</p><p><b>CONCLUSIONS</b>5-Aza-CdR inhibits the growth of human colon cancer cells in culture and in nude mice, and induces the cancer cells to re-express CDH13 in nude mice. Its mechanism may be that demethylation of the methylated CDH13 promoter induced by 5-Aza-CdR restores CDH13 expression and thus inhibits the tumor growth in nude mice.</p>

Effect of 5-Aza-CdR on expression and methylation of E-cadherin gene in human colon carcinoma cells / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Colon cancer is one of the most common malignant tumors, and its pathogenesis is not fully understood. Transcriptional silencing by DNA methylation is believed to be an important mechanism of carcinogenesis. E-cadherin can suppress tumor cell invasion and metastasis, and is considered as an invasion/metastasis suppressor gene. Inactivation of E-cadherin gene often occurs in colon carcinoma. This study was to investigate the correlation between E-cadherin gene expression and the methylation status of E-cadherin 5' CpG islands in human colon carcinoma cell line HT-29, and to explore the mechanism of carcinogenesis of colon cancer.</p><p><b>METHODS</b>Immunocytochemical dicho-step method and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expression of E-cadherin protein and mRNA in HT-29 cells after 5-Aza-CdR treatment; methylation specific PCR was used to analyze the methylation status at promoter of E-cadherin gene.</p><p><b>RESULTS</b>The expression of E-cadherin gene could be restored by 5-Aza-CdR treatment, immunocytochemical staining showed the positive expression ratio of E-cadherin increased from (21+/-7)% (1 micromol/L) to (39+/-13)% (5 micromol/L); E-cadherin genes were methylated and not expressed in HT-29 cells in the colon carcinoma.</p><p><b>CONCLUSIONS</b>E-cadherin methylation plays an important role in the carcinogenesis of colon carcinoma cells and can re-express after the treatment with 5-Aza-CdR.</p>