ING4 is negatively correlated with microvessel density in colon cancer.

ING4 is a novel tumor suppressor which is downregulated in a number of cancers. In this study, we investigated the role of ING4 in tumor angiogenesis in colorectal carcinoma (CRC) patients. Semi-quantitative RT-PCR, western blots, and immunohistochemistry were used to determine ING4 mRNA and protein expression in CRC and normal tissue from 60 CRC specimens and 30 colonic adenoma specimens. The correlation between ING4 expression and clinical stage, histological grade as well as lymph node metastasis was evaluated. Immunohistochemistry was performed to explore the correlation between ING4 expression and microvessel density (MVD) in CRC. CRC tissue had significantly lower levels of ING4 mRNA and protein compared to colonic adenoma and normal intestinal tissue. Immunostaining showed ING4 expression in 38 (63.3 %), 30 (100 %), and 60 (100 %) cases of normal colonic mucosa, adenoma, and normal intestinal mucosal tissue, respectively. Lower ING4 levels correlated with higher clinical stage and histological grade. ING4 mRNA and protein levels were significantly lower in CRC patients with lymph node metastasis compared to patients without lymph node metastasis (0.41 ± 0.30 vs. 0.91 ± 0.29 and 0.60 ± 0.21 vs. 0.87 ± 0.27, respectively; p < 0.001). Importantly, ING4 mRNA and protein levels were negatively correlated with MVD in CRC patients (p < 0.001). Our data suggest that ING4 levels are a potential biomarker of CRC progression and that ING4 may inhibit tumor growth by modulating angiogenesis in CRC.

Knock-down of ubiquitin-specific protease 22 by micro-RNA interference inhibits colorectal cancer growth.

PURPOSE: Increasing experimental evidences suggest that ubiquitin-specific protease 22 (USP22), a cancer stem cell marker, plays a crucial role in pathological processes of epithelial malignancies and other solid tumors, which makes it a potential target for cancer therapy. The aim of this study was to study the roles of USP22 in human colorectal cancer cell line HCT116 by suppressing USP22 expression with micro-interfering RNA (miRNA). METHODS: With the knock-down of USP22, the changes of cellular proliferation, cell cycle, cell apoptosis, and major vault protein (MVP) expression were investigated. Furthermore, a tumor xenograft model in nude mice was injected with USP22 miRNA silencing vector and the immunohistochemical staining was performed to evaluate the USP22 expression in the tumor. RESULTS: The knock-down of USP22 protein expression by miRNA resulted in the inhibition of cellular proliferation, the accumulation of cells in the G1 phase, the reduction of apoptosis, and the down-regulation of MVP expression. Furthermore, with orthotopic mice as a model, tumor growth was suppressed when USP22 miRNA silencing vector was injected. Immunohistochemical analyses of tumor sections revealed that USP22 expression in animals decreased when USP22 expression was inhibited by miRNA. CONCLUSION: These results support the hypothesis that USP22 plays a crucial role in tumor formation and growth by regulating cell proliferation with USP22-dependent signaling pathway. Furthermore, USP22 acts as a major transcriptional factor to regulate MVP drug resistant gene. Taken together, targeting USP22 may offer additional possibilities in cancer therapy.

Aberrant expression of USP22 is associated with liver metastasis and poor prognosis of colorectal cancer.

BACKGROUND: The present study was aimed at clarifying the expression of ubiquitin carboxyl-terminal hydrolase 22 (USP22), a novel deubiquitinating enzyme gene, in colorectal cancer (CRC) and its clinical significance. METHODS: USP22 expression was detected with quantitative RT-PCR, Western blot, and immunohistochemistry (IHC) in 43 CRCs and non-cancerous matched tissues. Furthermore, USP22 protein expression was analyzed in 192 CRC tumors by IHC to evaluate the association with survival. RESULTS: In 43 paired fresh tissues, the expression level of USP22 was significantly higher in primary CRCs than that in the paired non-cancerous tissues at both mRNA and protein levels (P < 0.0001). Nuclear USP22 expression significantly increased from normal mucosa through adenoma to primary carcinoma (P < 0.0001) and from primary carcinoma to liver metastasis (P = 0.021). The incidence of positive USP22 expression was 54.16% in 192 conventional CRC tissues. Notably, high USP22 expression was significantly associated with shorter disease-specific survival (P < 0.0001) and shorter disease-free survival (P < 0.0001). Cox regression analysis showed USP22 was an independent prognostic parameter for CRC patients. CONCLUSION: USP22 might be an independent predictive factor for CRC prognosis and aberrant expression of USP22 may play an essential role in colorectal carcinogenesis and liver metastasis.

Increased expression of ubiquitin-specific protease 22 can promote cancer progression and predict therapy failure in human colorectal cancer.

BACKGROUND AND AIMS: Increasing experimental evidence suggests that ubiquitin-specific protease 22 (USP22) could exhibit a critical function in pathological processes, including oncogenesis and cell cycle progression. The aim of this study was to investigate the role of USP22 and the association with its potential targets in colorectal cancer (CRC). METHODS: We evaluated the implication of USP22 and the candidate targets, such as B-cell-specific murine leukemia virus integration site-1 (BMI-1), cellular homolog of avian myelocytomatosis virus oncogene (c-Myc), cyclin D2, inhibitor of cyclin-dependent kinase (CDK) 4 (p16INK4a), and an alternate reading frame product of the CDKN2A locus (p14ARF), in matched samples comprising carcinoma and adjacent non-cancerous mucosa from 82 patients with CRC using quantitative reverse transcription-polymerase chain reaction and immunostaining analyses. RESULTS: The USP22 mRNA expression in the CRC tissues was significantly higher than those in the non-cancerous mucosa tissues (P < 0.0001). Increased mRNA expression of USP22 was associated with advanced American Joint Committee on Cancer stage (P = 0.033) and high likelihood of therapy failure after radical resection (P < 0.0001). The Cox regression analysis revealed that the USP22 mRNA expression level was a significant factor for predicting prognosis (P < 0.0001). The statistical correlation analysis in mRNA levels showed that USP22 was strongly correlated with BMI-1 (r = 0.790, P < 0.0001), c-Myc (r = 0.528, P < 0.0001), and cyclin D2 (r = 0.657, P < 0.0001), but not p16INK4a (r = 0.103, P = 0.358) or p14ARF (r = -0.039, P = 0.731). CONCLUSION: Our results indicate that activation of USP22 correlates with CRC progression and therapy failure. Additionally, the oncogenic role of USP22 in the progression of CRC can be mechanistically linked with BMI-1, c-Myc, and cyclin D2, but not with p16INK4a and p14ARF.

[Inhibitory effect of resveratrol on the growth of human colon cancer ls174t cells and its subcutaneously transplanted tumor in nude mice and the mechanism of action].

OBJECTIVE: To explore the effect and mechanism of resveratrol against human colon cancer ls174t cells in vitro and the growth of colon cancer in tumor-bearing nude mice. METHODS: MTT method was used to test the inhibiting effect of resveratrol on the growth and proliferation of ls174t cells. Transmission electron microscopy was used to observe the morphological changes of cell apoptosis, and FCM assay was performed to measure the changes of cell apoptosis rate and cell cycle. RT-PCR method was used to detect the expression of bcl-2 and bax mRNA, and Western blot was used to detect the expression of bcl-2 and bax protein. RESULTS: MTT test revealed that resveratrol showed significant inhibiting effect on ls174t cells in a concentration- and time-dependent manner. In the concentration range of 25, 50, 100, 200 and 400 micromol/L, the inhibition rate after resveratrol treatment for 24 hours was respectively 1.0%, 9.1%, 17.4%, 27.8% and 66.5%, while the inhibition rate after treatment for 48 hours was respectively 3.6%, 13.7%, 30.2%, 58.4% and 86.1%, and the inhibition rate after treatment for 72 hours was 18.1%, 33.0%, 48.6%, 61.2% and 89.4%, respectively, showing a very significant difference (P < 0.01). Typical ultrastructural apoptotic changes were observed in resveratrol-treated ls174t cells. It was found through FCM assay that resveratrol caused apoptosis in ls174 cells and blocked the cell cycle at S phase. RT-PCR and Western blot test showed that after the treatment of colon cancer cells with resveratrol at different concentrations (25, 50, 100 and 200 micromol/L), the expression level of bcl-2 was decreased, while expression level of bax was increased. The highest inhibition rate was 47.9%. In 200 mg/kg and 800 mg/kg resveratrol treatment groups, the weight of subcutaneously transplanted tumors in nude mice was 4.10 +/- 0.18 g and 3.05 +/- 0.35 g, respectively, the difference was significant compared with that of the control group (P < 0.01). CONCLUSION: Resveratrol can inhibit the growth of ls174t cells through apoptosis induction. The mechanism is probably related to inhibition of anti-apoptotic factor bcl-2 and enhancement of expression of apoptotic factor bax.

[Exploration on the modification of macroscopic classification of colorectal cancer].

OBJECTIVE: To explore the feasibility and clinical significance of a modified macroscopic classification of colorectal cancer. METHODS: The data of 1379 patients with colorectal cancer surgically treated between 1975 and 2003 were retrospectively analyzed. The patients were divided into four groups according to the primary macroscopic appearance: protruding type (group 1), local ulcerative type (group 2), invasive type (group 3) and non-invasive ulcerative type (group 4). The new classification system was simplified into two types: non-invasive type (group A, including group 1 and 2) and invasive type (group B, including group 3 and 4). The histo-differentiation, invasive depth into the intestinal wall, distance and number of lymph node metastasis and 5-year survival rate were analyzed and compared among the groups. RESULTS: There was no significant difference between group 1 and 2, and between group 3 and 4 in histodifferentiation, invasive depth into the intestinal wall, distance and number of lymph node metastasis and 5-year survival rate (P>0.05). However, after modification of the primary macroscopic classification, a significant difference was observed in all the above mentioned parameters between group A and group B (P<0.05). CONCLUSION: Our results demonstrate that the clinicopathological characteristics of the group 1 and 2, and of the group 3 and 4 are similar to each other. So it is reasonable to merge the protruding type and local ulcerative type into non-invasive type, while invasive type and non-invasive ulcerative type into invasive type. This simplified macroscopic classification should be practical and instructive in diagnosis, treatment and prognosis of colorectal cancer.

[Time pattern and prognostic evaluation of the recurrence of rectal cancer after resection].

OBJECTIVE: To investigate the regular pattern and prognostic evaluation of patients with recurrent rectal cancer after resection. METHODS: Three hundred and fourteen cases with recurrent rectal cancer after resection treated between May 1979 and November 2006 were classified into three groups according to the recurrence intervals (<3 years, 3-5 years, >5 years). The survival rates and prognosis in the three groups were analyzed and compared retrospectively. RESULTS: Of the 314 patients, the cancer relapsed in 247 cases (78.7%) in less than 3 years, and the recurrence occurred predominantly within 2 years (207 cases, 65.9%) after the operation. The neoplasm in 41 cases (13.3%) recurred in 3-5 years after the operation, and 26 cases (8. 3%) in more than 5 years after the resection. Disease-free interval, Dukes stage, neoplasm gross type, histological type, T stage, lymphatic and distant metastasis were associated with the prognosis on univariate analysis. And disease-free interval and tumor Dukes stage were independent prognostic factors for survival rate on multivariate analysis. Disease-free interval and progression-free survival were related positively with survival time. CONCLUSIONS: The rectal cancer patients should be followed-up intensively for 2 years after the operation and moderately from then on. Disease-free interval and progression-free survival could be taken as the best predictors of long-term cure and prognosis.

Effectiveness of rehabilitation training combined with acupuncture on aphasia after cerebral hemorrhage: A systematic review protocol of randomized controlled trial.

BACKGROUND: This study aims to systematically evaluate the effectiveness of rehabilitation training (RT) combined with acupuncture on aphasia after cerebral hemorrhage (CH). METHODS: PUBMED, Cochrane Central Register of Controlled Trials, EMBASE, Web of Science, Ovid, Cumulative Index to Nursing and Allied Health Literature, Allied and Complementary Medicine Database, Chinese Biomedical Literature Database, and China National Knowledge Infrastructure will be searched to identify any potential studies from inception to March 1, 2019, without language restrictions. All randomized controlled trials and case-controlled studies assessing the effectiveness of RT combined with acupuncture for the treatment of aphasia following CH will be included in this study. Cochrane risk of bias tool will be used to determine the methodological quality for included studies. RevMan 5.3 software (Cochrane Community, London, UK) will be utilized to perform statistical analysis. RESULTS: This study will systematically evaluate the effectiveness of RT and acupuncture for aphasia post CH. Primary outcome includes aphasia, which can be measured by Aachener Aphasia Test or Communicative Activity Log or other related scales. Secondary outcomes consist of speech performance, as assessed by Western Aphasia Battery-Revised; measure of skill in Supported Conversation scales; measure of Participation in Conversation scales; types of strategies used in conversation; occurrence and repair of conversation breakdowns; as well as any adverse events. CONCLUSION: The results of this study will provide present evidence on assessing effectiveness of RT and acupuncture after CH. DISSEMINATION AND ETHICS: The findings of this study are expected to be published in peer-reviewed journals. It does not require ethical approval, because no individual data will be utilized in this study. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42019131587.

[The effect of As2O3 on induction of apoptosis and inhibition of telomerase activity in colon cancer LS-174T cells].

OBJECTIVE: To study the impact of arsenic trioxide (As2O3) on human colorectal carcinoma LS-174T cells and their activity of telomerase. METHODS: LS-174T cells and xenograft model of nude mice were treated with As2O3. The inhibitory effect of As2O3 on survival of LS-174T cells was determined by MTT assay. Apoptosis was determined by electron microscopy and fluorescence microscopy. Cell cycle was assessed by flow cytometry. Telomerase activity in LS-174T cells was determined by PCR-ELISA kit. RESULTS: With the increasing concentration of As2O3, the ratio of living cells to dead cells decreased significantly, and the IC50 value was 5.23 micromol/L. Apoptosis curve appeared after 24 h and cells turned to apoptosis in a time-dependent manner. As2O3 inhibited the telomerase activity in cell extraction, obviously in a concentration-dependent and time-dependent manner. Inhibitiory effect of As2O3 on xenograft model of nude mice was observed by tumor volume and weight measurement, showing a significant difference between As2O3 and control groups (P < 0.05). CONCLUSION: Both the experiments in vitro and in vivo showed an inhibitory effect of As2O3 on colonrectal cancer S-174T cell growth, probably by induction of apoptosis and inhibition of telomerase activity.

[Effect of lateral lymph nodes dissection and autonomic nerve preservation in anterior resection for rectal cancer: 124 cases review].

OBJECTIVE: To investigate the clinical effect of lateral lymph nodes dissection and autonomic nerve preservation in anterior resection for rectal cancer. METHODS: One hundred and twenty-four patients with rectal cancer underwent anterior resection with lateral lymph nodes resection and autonomic nerve preservation. The patients were followed-up through post-operational questionnaire about the function of defecation, urination and sex after the operation. And post-operative survival was analyzed retrospectively. RESULTS: Urinary catheters were removed in 112 cases (90.3%) in 3 days post operation, the mean time of indwelling catheter was (58.3 +/- 2.1) h. Nineteen patients experienced fecal incontinence, 12 cases of them recovered through release-training and one recovered spontaneously. Of the 98 questionnaire respondents, 61 cases (62.3%) could erect normally, and 56 cases (57.1%) had normal sexual function. The max-micturition-desire urine volume was (401.2 +/- 23.1) ml and the residual urine volume was (28.2 +/- 2.2) ml. Five year survival rate of all the patients was 61.2%. CONCLUSIONS: Lateral lymph nodes dissection and autonomic nerve preservation in anterior resection for rectal cancer can decrease the post-operative dysfunction of defecation, urination and sex life and does not affect the survival.

Abnormal expression of p190RhoGAP in colorectal cancer patients with poor survival.

PURPOSE: This study aims to investigate the expression and clinical significance of p190RhoGAP, a member of the RhoGAP family, in colorectal cancer (CRC). METHODS: The expression p190RhoGAP was detected by RT-PCR, western blot (WB) and immunohistochemistry (IHC) in 14 paired CRCs and matched non-cancerous mucosal tissues. The protein content of p190RhoGAP was identified in 114 CRCs by IHC. In addition, the association of the expression of p190RhoGAP with carcinogenesis, distant metastasis and prognosis was further evaluated. RESULTS: In 14 paired fresh tissues, the mRNA (P<0.0001) and protein (P = 0.003) expression levels of p190RhoGAP were significantly higher in primary CRCs than in paired non-cancerous mucosal tissues; and was consistent with WB results. The expression of p190RhoGAP increased from normal mucosa to adenoma, and became even greater in primary carcinoma (P = 0.001). The expression level of p190RhoGAP was highest in liver metastasis compared to primary carcinoma (P = 0.028). The incidence of p190RhoGAP expression-positive cases was 58.77% in 114 CRC tissues. Furthermore, the enhanced expression of p190RhoGAP was significantly associated with shorter disease-specific survival (P<0.001) and shorter disease-free survival (P<0.001). Cox regression analysis indicated that p190RhoGAP was an independent prognostic parameter for CRC. CONCLUSION: p190RhoGAP may be an independent predictive factor for the prognosis of CRC, and the abnormal expression of p190RhoGAP may play a crucial role in colorectal carcinogenesis and distant metastasis.

Effect of a nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester on invasion of human colorectal cancer cell line SL-174T.

AIM: To investigate the effect and mechanism of action of the nitric oxide synthase (NOS) inhibitor NG-nitro-L-arginine methyl ester (L-NAME) on invasion and metastasis of human colorectal cancer cell line SL-174T. METHODS: Human colorectal cancer cell line SL-174T was cultured and treated separately with four different dosages of L-NAME for 72 h. Nitric oxide (NO) production was measured with Griess reagent. The effect of L-NAME on invasion and migration of SL-174T cells were evaluated by using Transwell chambers attached with polycarbonate filters and reconstituted basement membrane (Matrigel). RT-PCR was performed to determine the mRNA levels of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor metalloproteinase-2 (TIMP-2). RESULTS: L-NAME could significantly inhibit NO production of SL-174T in a dose-dependent manner. After being treated for 72 h with 0.2, 0.4, 0.8, and 1.0 mmol/L L-NAME, respectively, the ability of the L-NAME treated SL-174T cells to invade the reconstituted basement membrane decreased significantly (t = 8.056, P< 0.05; t = 14.467, P< 0.01; t = 27.785, P< 0.01; and t = 29.405, P< 0.01, respectively) and the inhibition rates were 10.29%, 19.62%, 34.08%, and 42.23%, respectively. Moreover, L-NAME could inhibit migration of SL-174T cells, and the inhibition rates were 20.76%, 24.95%, 39.43%, and 46.85% for L-NAME at 0.2, 0.4, 0.8, and 1.0 mmol/L, respectively (t= 15.116, P< 0.01). In addition, after treatment with L-NAME, expression of MMP-2 mRNA was significantly decreased (t = 71.238, P< 0.01) and that of TIMP-2 mRNA was markedly increased (t = -13.020, P< 0.01). CONCLUSION: L-NAME exerts anti-invasive and anti-metastatic effects on SL-174T cell line via downregulating MMP-2 mRNA expression and upregulating TIMP-2 mRNA expression.

[Radiosensitivity on colorectal neoplasms by recombinant adenoviral-mediated wild-type p53 gene].

OBJECTIVE: To study the radiosensitization on the cells of colorectal cancer transfected with recombinant adenovirus vector-mediated wild-type p53. METHODS: SW480 cells transfected by wild-type p53 were treated with 4 Gy and 6 Gy radiation. The expression of recombinant adenovirus vector-mediated wild-type p53 gene was detected by Western blotting. The inhibition rate of SW480 cells was examined by MTT, apoptotic rate by TdT-mediated dUTP nick end labeling (TUNEL) and proliferating cell nuclear antigen (PCNA) by immunohistochemical method. RESULTS: SW480 cells transfected by wild-type p53 were inhibited significantly by 4 Gy and 6 Gy radiation. The level of apoptosis increased and the expression of PCNA decreased. CONCLUSION: Cells of colorectal carcinoma transfected with wild-type p53 increases their radiation sensitivity.

The potential of plasma miRNAs for diagnosis and risk estimation of colorectal cancer.

Circulating microRNAs (miRNAs) were recognized to be potential non-invasive biomarkers for colorectal cancer (CRC) detection and prediction. Meanwhile, the association of the expression of plasma miRNAs with the risk of CRC patients has rarely been analyzed. Therefore, we conducted this study to evaluate the value of plasma miRNAs for CRC diagnosis and risk estimation. Fasting blood samples from 100 CRC patients and 79 cancer-free controls were collected. Plasma miR-106a, miR-20a, miR-27b, miR-92a and miR-29a levels were detected by RT-qPCR. Sensitivity and specificity were employed to evaluate the diagnostic value of miRNAs for CRC. Univariate and multivariate logistic regression were employed to analyze the association between miRNAs expression and CRC risk. As results, miR-106a and miR-20a were elevated in the patients with CRC. The sensitivity of miR-106a was 74.00% and the specificity was 44.40%, while the cutoff value was 2.03. As for miR-20a, the sensitivity was 46.00% and specificity was 73.42% when employed 2.44 as cutoff value. High expression of plasma miR-106a increased CRC risk by 1.80 -fold. Plasma miR-106a and miR-20a may as noninvasive biomarkers for detecting the CRC. High expression of miR-106a associated with CRC risk.

Effects of endostatin on expression of vascular endothelial growth factor and its receptors and neovascularization in colonic carcinoma implanted in nude mice.

AIM: To investigate the antiangiogenic effects of endostatin on colonic carcinoma cell line implanted in nude mice and its mechanism. METHODS: Nude mice underwent subcutaneous injection with LS-174t colonic carcinoma cell line to generate carcinoma and were randomly separated into two groups. Mice received injection of vehicle or endostatin every day for two weeks. After the tumor was harvested, the tumor volumes were determined, and the expressions of CD34, VEGF and Flk-1 were examined by immunohistochemical method. RESULTS: Tumor volume was significantly inhibited in the endostatin group (84.17%) and tumor weight was significantly inhibited in the endostatin group (0.197+/-0.049) compared to the control group (1.198+/-0.105) (F = 22.56, P = 0.001), microvessel density (MVD) was significantly decreased in the treated group (31.857+/-3.515) compared to the control group (100.143+/-4.290) (F = 151.62, P<0.001). Furthermore, the expression of Flk-1 was significantly inhibited in the treated group (34.29%) compared to the control group (8.57%) (chi(2) = 13.745, P = 0.001). However no significant decrease was observed in the expression of vascular endothelial growth factor (VEGF) between these two groups (chi(2) = 0.119,P = 0.730). CONCLUSION: Endostatin can inhibit tumor growth and angiogenesis by blocking Vegf/Flk-1 pathway. This experiment provides the theory basis for developing a new anti-carcinoma drug through studying the properties of anti-angiogenesis inhibitors.

Reversal of 5-flouroucial resistance by adenovirus-mediated transfer of wild-type p53 gene in multidrug-resistant human colon carcinoma LoVo/5-FU cells.

AIM: To observe the reversal effects of wide-type p53 gene on multi-drug resistance to 5-FU (LOVO/5-FU). METHODS: After treatment with Ad-p53, LOVO/5-FU sensitivity to 5-Fu was investigated using tetrazolium dye assay. Multidrug resistance gene-1 (MDR1) gene expression was assayed by semi-quantitative reverse transcription-polymerase chain reaction and the expression of p53 protein was examined by Western blotting. RESULTS: The reversal activity after treatment with wide-type p53 gene was increased up to 4.982 fold at 48 h. The expression of MDR1 gene decreased significantly after treatment with wide-type p53 gene, and the expression of p53 protein lasted for about 5 d, with a peak at 48 h, and began to decrease at 72 h. CONCLUSION: Wide-type p53 gene has a remarkable reversal activity for the high expression of MDR1 gene in colorectal cancers. The reversal effects seem to be in a time dependent manner. It might have good prospects in clinical application.

[On rationality of total mesorectal excision for rectal cancer].

OBJECTIVE: To discuss the rationality and clinical significance of total mesorectal excision (TME) assessed by the pattern of local recurrence of rectal cancer. METHODS: 207 rectal cancer patients treated by surgery from 1975 to 2001 who developed local recurrence were retrospectively studied as to the option of treatment based on their outcome. 172 of these 207 patients had complete record of follow-up with a follow-up rate of 83.0%. RESULTS: The local recurrent sites of 207 patients were: anastomotic stoma 71, mesorectum 65, perineum 50, lymph nodes 59, multiple recurrences 89 and others sites rare. Among 81 patients who had been first treated by anterior resection (AR), 58 were again treated by abdomino-perineal resection (APR). Among 102 patients who had been treated by APR, 38 were treated by perineal tumor resection, 15 were treated by total and posterior pelvic organ resection, and 18 were supplemented with lymph node resection. By all modalities, the resection rate was 66.1% (137/207), in which the curable resection rate was 46% (63/137). The 5-year survival rate of patients with tumor resection was 23.3% (32/137), those treated by curable resection was 34.9% (22/63). CONCLUSION: From the pattern of local recurrence, the operation for middle-lower rectal cancer must follow the principle of total mesorectum excision (TME). Patients with local recurrence should be energetically treated in the second operation if tolerable.

TFAP2E hypermethylation was associated with survival advantage in patients with colorectal cancer.

PURPOSE: Hypermethylation of TFAP2E (AP-2E) is associated with the chemotherapy-resistant in patients with colorectal cancer (CRC), but its implications on prognosis directly remain unknown. This study was aimed to investigate the role of AP-2E methylation status and other clinicopathologic parameters as predictors of prognosis. METHODS: We detected the methylation status of AP-2E in tumor and adjacent non-tumor tissues from 311 sporadic CRC patients by methylation-sensitive high-resolution melting analysis. Log-rank tests and multivariate Cox analyses were performed to evaluate the role of AP-2E methylation status and other clinicopathologic parameters as predictors of prognosis. RESULTS: Hypermethylation of AP-2E was detected in 61 % (190/311) tumor tissues. It occurred more frequently in tumors in earlier stages (I/II; P = 0.02), lower levels of tumor invasion (T1-T3; P = 0.04), fewer lymph nodes involved (N0; P < 0.01), and higher histologic grades (G1/G2; P < 0.01). The overall 5-year survival rates in hypermethylation and hypomethylation group were 76.91 and 47.17 % (P < 0.0001), respectively. AP-2E hypermethylation was significantly associated with a favorable clinical outcome with a hazard ratio of 0.486 (95 % CI 0.342-0.692, P < 0.0001) after controlling for age, gender, tumor location, histologic type, TNM staging, and histologic grade. CONCLUSIONS: AP-2E was frequently hypermethylated in tumors from patients with CRC. Aberrant hypermethylation of AP-2E occurred more frequently in tumors with earlier stages, lower levels of tumor invasion, fewer lymph nodes involved, and higher histologic grades. AP-2E hypermethylation might be an independent predictor of survival advantage in patients with CRC.

Induction of apoptosis by D-limonene is mediated by inactivation of Akt in LS174T human colon cancer cells.

D-limonene is recognized as a potential chemotherapeutic agent, however, the details of this mechanism remain unclear. In this study, we investigated the effects of d-limonene on colon cancer cell viability and its potential mechanism of action in vitro. After 48 h of treatment, d-limonene suppressed the viability of LS174T cells in a dose-dependent manner and caused a dose-dependent apoptotic cell death. D-limonene activated caspase-3 and -9 and PARP cleavage in a dose-dependent manner. Moreover, an increase in Bax protein and cytosol cytochrome c from mitochondria and a decrease in bcl-2 protein were observed following treatment with d-limonene. In addition, d-limonene decreased the levels of p-Akt (Ser473), p-Akt (Thr308) and p-GSK-3ß (Ser9), suggesting that d-limonene induced apoptosis via the mitochondrial death pathway and the suppression of the PI3K/Akt pathway.