ABSTRACT
Colorectal cancer (CRC) is the third most commonly diagnosed cancer. Celastrol exhibits anti-tumor activities in a variety of cancers. However, the effect of Celastrol on human CRC and the underlying mechanisms still need to be elucidated. The present study aimed to use in vitro and in vivo methods to clarify the anti-tumor effect of Celastrol and use protein microarrays to explore its mechanisms. We demonstrated that Celastrol effectively inhibited SW480 CRC cell proliferation. Two weeks of Celastrol gavage significantly inhibited the growth of xenografts in nude mice. A total of 69 candidate proteins were identified in the protein microarray experiment, including the most highly enriched protein Shoc2, which is a scaffold protein that modulates cell motility and metastasis through the ERK pathway. Celastrol significantly inhibited ERK1/2 phosphorylation in cell lines and xenograft tumors. Down-regulation of Shoc2 expression using Shoc2 siRNA also inhibited ERK1/2 phosphorylation. Furthermore, down-regulation of Shoc2 expression also significantly inhibited proliferation, colony formation, and migration functions of tumor cells. In addition, the LD0 of Celastrol by gavage is equal or more than 80 mg/kg in C57 male mice. In summary, we unraveled the anti-CRC function of Celastrol and confirmed for the first time that it inhibited the ERK1/2 pathway through binding to Shoc2.
Subject(s)
Cell Proliferation/drug effects , Colorectal Neoplasms/drug therapy , Intracellular Signaling Peptides and Proteins/genetics , Triterpenes/administration & dosage , Animals , Colorectal Neoplasms/genetics , Colorectal Neoplasms/pathology , Gene Expression Regulation, Neoplastic/drug effects , Humans , MAP Kinase Signaling System/drug effects , Mice , Pentacyclic Triterpenes , Protein Array Analysis , Protein Binding/genetics , RNA, Small Interfering/genetics , Xenograft Model Antitumor AssaysABSTRACT
<p><b>OBJECTIVE</b>To evaluate association between DNA methylation of MAL, CDKN2A, and MGMT in stool and development of colorectal cancer, and to evaluate the screening value of these biomarkers in colorectal cancer and pre-malignant lesions.</p><p><b>METHODS</b>Morning stool specimens were collected from 69 patients with colorectal cancer, 24 with colon adenoma, 19 with hyperplastic polyps, and 26 healthy controls. DNA was extracted and treated with bisulfite. Methylation-specific PCR(MSP) was performed for methylation analysis of MAL, CDKN2A and MGMT in DNA samples. Associations between clinicopathological features and gene methylation were analyzed. The sensitivity of diagnosis by combining three methylation markers was compared with fecal occult blood test(FOBT).</p><p><b>RESULTS</b>The methylation frequencies of MAL, CDKN2A and MGMT were 78.3%, 52.5% and 55.1% in colorectal cancer, 58.3%, 41.7% and 37.5% in colon adenomas, 26.3%, 15.8% and 10.5% in hyperplastic polyps, and 3.8%, 0 and 3.8% in healthy controls, respectively. Significant differences in three genes were found between colorectal cancer and hyperplastic polyp, colorectal cancer and healthy control, colon adenoma and hyperplastic polyp, colon adenoma and healthy control(all P<0.05). The diagnostic sensitivity by combining three methylation markers was 92.8% in colorectal cancer, 70.8% in colon adenomas, significantly higher than FOBT examination (29.0% in colorectal cancer and 25.0% in colon adenomas, all P<0.05). No significant associations existed between three genes methylation of the three genes and clinical characteristic including sex, age, tumor location, lymph node metastases and TNM stage (all P>0.05).</p><p><b>CONCLUSION</b>DNA methylations levels of MAL, CDKN2A, and MGMT in stools are significantly higher in colorectal cancer and colon adenoma, which may serve as an noninvasive approach for the screening of colorectal cancer and pre-malignant lesions.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Colorectal Neoplasms , Diagnosis , Genetics , Cyclin-Dependent Kinase Inhibitor p16 , Genetics , DNA Methylation , Early Detection of Cancer , Feces , Chemistry , Mass Screening , O(6)-Methylguanine-DNA Methyltransferase , Genetics , Precancerous Conditions , Diagnosis , Genetics , Promoter Regions, Genetic , GeneticsABSTRACT
Objective: To understand the psychological status of the rescue troops one month after rescue action for the devastating earthquake and to explore the effect of some important events on the psychological status of soldiers. Methods: The subjects were randomly selected from the 3 major rescue forces, a Cannon troop from PLA Chengdu Military Area Command, a Cannon troop from PLA Jinan Military Area Command and Armed Police Forces of Sichuan Province, 32-42 days after the disaster rescue. A total of 170 subjects were tested with Symptom Checklist-90(SCL-90), and the results were compared with the norms of normal Chinese and Chinese serviceman. Student's t test was used to determine the significance of the differences. Results: A total of 78 valid questionnaires were analyzed. The total score of SCL-90 was 130. 25 ± 33. 07 for the rescuers, the positive items was 27.59 ± 15.75, and the mean factorial score was 1.28-1.64. The somatization score was higher (P<0.05) and the interpersonal sensitivity score was lower (P<0.01) than the norms of the Chinese. The factorial scores of obsession, interpersonal sensitivity, depression, anxiety, paranoid and psychotic were lower than the norm of Chinese servicemen (all P< 0.01). Soldiers with experience of searching dead bodies had lower scores in sleep and food consumption, and higher score of positive items (all P<0.01 for each) compared those without the related experience. Conclusion: The psychological status of the earthquake-rescuers is basically normal 32-42 days after the earthquake. Some positive items are associated with the experience such as body treatments. Surveillance of psychological disorders in these troops is helpful for the early diagnosis and intervention of psychological problems.
ABSTRACT
Objective:To investigate frequencies of microsatellite instability(MSI)and loss of heterozygosity(LOH)in renal ceil carcinoma(RCC),and to discuss the relationship of clinicopathological characteristics of RCC with MSI and LOH. Methods:Twelve microsatellite markers located at chromosomes 3p,9p and 14q were selected to investigate microsatellite alterations(MSI and LOH)in 31 RCC specimens and their paired metastasis specimens by polymerase chain reaction- polyacrylamide gel elect rophoresis-ethylene dibromide(PCR-PAGE-EB)staining and sequencing.Results:The frequency of MSI could reached 61.3% and that of LOH could reach 54.8%.The highest frequency of MSI was at locus of D9S168(32.3%);the highest frequency of LOH was at locus of D3S1289(21.4%).No correlation was found between MSI or LOH and the patients' age,sex,pathology type and metastastis,except that MSI was correlated with TNM stage of RCC(P
