Sulforaphane suppresses carcinogenesis of colorectal cancer through the ERK/Nrf2­UDP glucuronosyltransferase 1A metabolic axis activation.

The long pre­cancerous state of colorectal cancer (CRC) provides an opportunity to prevent the occurrence and development of CRC. The detoxification of CRC food­borne carcinogenic heterocyclic amines is highly dependent on UDP glucuronosyltransferase 1A (UGT1A)­mediated glucuronidation. Sulforaphane (SFN), a phytochemical, possesses antioxidant, anti­inflammatory and anticarcinogenic effects on the prevention of CRC. Previous studies revealed that SFN upregulates the expression of UGT1A. The aim of the present study was to investigate the regulatory mechanism of SFN­induced UGT1A upregulation and provide novel understanding on the basic research and chemoprevention of CRC. In the present study, the viability and proliferation of CRC cells (HT­29 and SW480) treated with SFN were assessed by MTT, colony formation and EdU assays. Flow cytometry was used to detect the cell cycle arrest and apoptosis of cells treated with different concentrations of SFN. The motility of cells was determined by wound healing and Transwell assays. Nuclear factor, erythroid 2 like 2 (Nrf2) short hairpin RNA (shRNA) and negative control shRNA lentiviruses were used for cell transfection. Reverse transcription­quantitative polymerase chain reaction and western blotting were employed to verify the role of Nrf2 in SFN­induced UGT1A. HT­29 and SW480 cells were divided into a control, an SFN and a PD98059 [an extracellular signal­regulated kinase (ERK) inhibitor] + SFN group. Western blotting detected the protein levels of Nrf2 and UGT1A. Intracellular levels of reactive oxygen species (ROS) were detected using a reactive oxygen assay kit. The results revealed that SFN inhibits cell proliferation and colony formation, promotes apoptosis, and reduces the migratory ability of CRC cells. The phosphorylation of ERK induced by SFN promoted Nrf2 accumulation. Furthermore, a significant increase in the levels of UGT1A was observed, which coincided with SFN­induced upregulation of Nrf2 levels in nuclear fractions. Pretreatment with PD58059 reversed the SFN­induced subcellular translocation of Nrf2 and the expression of UGT1A. In addition, SFN­induced high levels of ROS in CRC cells may be associated with the ERK signaling pathway. Collectively, these results indicated that SFN inhibited the proliferation of CRC cells and upregulated the expression of UGT1A in CRC cells via the ERK/Nrf2 signaling pathway.

Screening and application of nutritional support in elderly hospitalized patients of a tertiary care hospital in China.

BACKGROUND: Malnutrition is very common in elderly patients admitted to the hospital. The aim of our study is to assess the nutritional status of elderly patients and the use of nutritional support in a tertiary care hospital in China and to analyze the impacts of nutritional status and nutritional support on clinical outcomes. METHODS: Statistical analysis was performed on a sample of 745 elderly patients in the geriatric medicine department of Qilu Hospital of Shandong University from March 2012 to March 2015. The Nutrition Risk Screening 2002 (NRS 2002) and Mini Nutritional Assessment-short forms (MNA-SF) were utilized for the nutritional risk screening at admission. Personal information, anthropometric measurements, laboratory tests, nutritional support and clinical outcomes were recorded. Comparisons were carried out to analyze impacts on clinical outcomes and prognosis based on incidence rate of nutritional risk, nutritional support rate, and different methods of support. RESULTS: NRS 2002 and MNA-SF were utilized to screen for nutritional risk at admission. The results of this screening were 39.81% and 44.10%, respectively. Based on the results of the MNA-SF, 33.38% of elderly patients were at risk of malnutrition and 5.5% were malnourished. The incidence of nutritional risk in the departments of Gastroenterology, Hematology, and Respiratory were 51.72%, 46.88%, 43.33%, respectively, higher than in other departments. Patients with nutritional risk were more likely to have a longer hospital stay compared to those without (P < 0.05). The nutritional support rate of patients overall was 16.49%, and the ratio of Parenteral nutrition (PN):Enteral nutrition (EN) was 5.13:1. Patients at nutritional risk had an in-hospital support rate of 29.63% and 28.57%, respectively, identified via screening by NRS 2002 and MNA-SF. Nutritional support rate of patients without nutritional risk was 7.8%(35/449) and 6.96%(29/417), respectively. Patients in the departments of Gastroenterology and Hematology had higher rates of nutritional support than patients in other departments. In addition, results showed that in patients with nutritional risk and malnutrition, nutritional support decreased the length of hospital stay (P<0.05). The patients that received nutritional support also had a lower incidence of infectious complications than the patients without nutritional support (NRS 2002 was 6.82%:18.18% and MNA-SF was 9.57%:20.23%)(P<0.05). CONCLUSIONS: Undernourishment and nutritional risk in elderly patients at hospital admission is a common occurrence. In the current study, the nutritional risk rate in the Gastroenterology department was higher than in other departments. Patients with normal nutritional status were still receiving nutritional support. Overall, there is a need to better apply nutritional support in the clinical treatment of elderly patients. In elderly patients with nutritional risk and malnutrition, nutritional support reduced the length of hospital stay and the incidence of infectious complications.

Research progress on chemopreventive effects of phytochemicals on colorectal cancer and their mechanisms.

Colorectal cancer (CRC) is a type of cancer with high morbidity and mortality rates worldwide and has become a global health problem. The conventional radiotherapy and chemotherapy regimen for CRC not only has a low cure rate but also causes side effects. Many studies have shown that adequate intake of fruits and vegetables in the diet may have a protective effect on CRC occurrence, possibly due to the special biological protective effect of the phytochemicals in these foods. Numerous in vitro and in vivo studies have demonstrated that phytochemicals play strong antioxidant, anti-inflammatory and anti-cancer roles by regulating specific signaling pathways and molecular markers to inhibit the occurrence and development of CRC. This review summarizes the progress on CRC prevention using the phytochemicals sulforaphane, curcumin and resveratrol, and elaborates on the specific underlying mechanisms. Thus, we believe that phytochemicals might provide a novel therapeutic approach for CRC prevention, but future clinical studies are needed to confirm the specific preventive effect of phytochemicals on cancer.

Correlation between Helicobacter pylori-associated gastric diseases and colorectal neoplasia.

AIM: To explore the correlation between Helicobacter pylori (H. pylori)-associated gastric diseases and colorectal neoplasia. METHODS: Patients included in this study underwent a colonoscopy and esophago-gastro-duodenoscopy (EGD) along with histopathological measurement between March 2012 and March 2015 at Qi-Lu Hospital of Shandong University, who also had results of H. pylori detection. A total of 233 cases were selected. Demographic data, H. pylori infection status (including results of rapid urease tests and gastric mucosa pathological examinations) and histopathological examination results of gastric and colorectal mucosa were gathered and analyzed. The statistical analysis focused on the prevalence of colorectal neoplasms among patients with various histopathological categories of the stomach. ORs and their 95%CI were calculated to describe the strengths of the associations. RESULTS: The incidence rates of colorectal adenoma without high-grade intraepithelial neoplasia (HGIEN) (OR = 2.400, 95%CI: 0.969-5.941), adenoma with HGIEN (5.333, 1.025-27.758) and adenocarcinoma (1.455, 0.382-5.543) were all higher for patients with H. pylori-associated gastritis than for those in the control group. The incidence rate of colorectal adenoma with HGIEN (3.218, 0.767-13.509) was higher in patients with intestinal metaplasia than in the control group, while the incidence rates of adenoma without HGIEN (0.874, 0.414-1.845) and adenocarcinoma (0.376, 0.096-1.470) were lower in the intestinal metaplasia group than in the control group. The incidence rate of colorectal adenoma without HGIEN (3.111, 1.248-7.753) was significantly higher in the gastric intraepithelial neoplasia group than in the control group, while the rates of adenoma with HGIEN (1.481, 0.138-15.941) and adenocarcinoma (2.020, 0.561-7.272) were higher in the gastric intraepithelial neoplasia group. Incidence rates of colorectal adenoma without HGIEN (1.067, 0.264-4.314), adenoma with HGIEN (2.667, 0.231-30.800) and adenocarcinoma (2.182, 0.450-10.585) were all higher in the gastric adenocarcinoma group than in the control group. CONCLUSION: H. pylori infection as well as H. pylori-associated gastric diseases are risk factors for colorectal neoplasia.

Effects of co-treatment with sulforaphane and autophagy modulators on uridine 5'-diphospho-glucuronosyltransferase 1A isoforms and cytochrome P450 3A4 expression in Caco-2 human colon cancer cells.

Sulforaphane (SFN), which is highly enriched in cruciferous vegetables, has been investigated for its cancer chemopreventive properties and ability to induce autophagy. Uridine 5'-diphospho (UDP)-glucuronosyltransferase (UGT)1A induction is one of the mechanisms that is responsible for the cancer chemopreventive activity of SFN. The current study demonstrates that rapamycin may enhance the chemopreventive effects of SFN on Caco-2 cells; this may be partially attributed to nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2)- and human pregnane X receptor (hPXR)-mediated UGT1A1, UGT1A8 and UGT1A10 induction. These results indicate that targeting autophagy modulation may be a promising strategy for increasing the chemopreventive effects of SFN in cases of colon cancer.

[Effects of autophagy modulator on autophagy and uridine 5'-diphospho-glucuronosyltransferase 1A1 induced by sulforaphane].

OBJECTIVE: To explore the effects of 3-methyladenine (3-MA) and rapamycin (Rapa) on autophagy and uridine 5'-diphospho-glucuronosyltransferase 1A1 (UGT1A1) induced by sulforaphane (SFN) in human colon cancer Caco-2 cells. METHODS: Western blot was used to detect the expression of microtubule-associated protein 1 light chain 3 (LC3) and UGT1A1 proteins. And immunocytochemistry was employed to observe the intracellular distribution of LC3 and nuclear localization of NF-E2-related factor 2 (Nrf2). Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) was employed to examine the mRNA expression of UGT1A1 and human pregnane X receptor (hPXR). RESULTS: After the treatment of SFN, the LC3-II protein was induced in a dose and time-dependent manner. SFN-induced LC3-II protein could be attenuated and enhanced by 3-MA and Rapa respectively. In comparison with the control group, UGT1A1 mRNA levels increased significantly after the treatment of Rapa, SFN or their combination (2.4, 4.12 and 2.41 folds respectively, all P < 0.01). And the combination of SFN and Rapa possessed the highest level. UGT1A1 protein band intensity was also enhanced in three groups. There was no obvious nuclear staining of Nrf2 in control group while intense nuclear fluorescent labeling of Nrf2 could be observed in the SFN-treated groups, especially the combination group of SFN and Rapa. The hPXR mRNA levels increased significantly in the Rapa and combination groups (1.82 and 1.4 folds respectively, both P < 0.01). CONCLUSION: The treatment of 3-MA or Rapa may attenuate or enhance SFN-induced autophagy respectively. And Rapa also potentiates SFN-induced UGT1A1 expression. The mechanism for the synergic effect of Rapa and SFN on UGT1A1 induction may be a simultaneous activation of Nrf2 and hPXR signaling pathway.

Polymorphic expression of UDP-glucuronosyltransferase UGTlA gene in human colorectal cancer.

BACKGROUND: Polymorphism of genes encoding drug-metabolizing enzymes is known to play an important role in increased susceptibility of colorectal cancer. UGT1A gene locus has been suggested to define tissue-specific glucuronidation activity. Reduced capacity of glucuronidation is correlated with the development of colorectal cancer. Therefore, we sought to explore polymorphism of UGTlA gene in human colorectal cancer. METHODS: Cancerous and healthy tissues were obtained from selectedpatients. Blood samples were collected and UGTlA mRNA transcriptions were analyzed. Genomic DNA was prepared and UGTlA8 exon-1 sequences were amplified, visualized and purified. The extracted DNA was subcloned and sequenced. Two-tailed Fisher's exact test, Odds ratios (ORs), confidence interval (CIs) and Logistics Regression Analysis were used for statistical analysis. RESULTS: UGTlA mRNA expression was reduced in cancerous tissues compared with healthy tissues from the same patient . The UGTlA mRNA expression of healthy tissue in study patients was lower than control . The mRNA expression of cancerous tissue was down-regulated in UGTlAl, 1A3, 1A4, lA6, 1A9 and up-regulated in UGTlA8 and UGTlAl0 UGT1A5 and UGT1A7 were not expressed in colonic tissue of either group. The allele frequency of WT UGTlA8*1 was higher (p = 0.000), frequency of UGTlA8*3 was lowered in control group (p = 0.000). The expression of homozygous UGTlA8*1 was higher in control group (p = 0.000). Higher frequency of both heterozygous UGTlA8*1/*3 and UGTlA8*2/*3 were found in study group (p = 0.000; p = 0.000). The occurrence of colorectal cancer was mainly related to the presence of polymorphic UGTlA8*3 alleles (p = 0.000). CONCLUSION: Regulation of human UGT1A genes is tissue-specific. Individual variation in polymorphic expressions of UGTlA gene locus was noted in all types of colonic tissue tested, whereas hepatic tissue expression was uniform. The high incidence of UGTlA8 polymorphism exists in colorectal cancer patients. UGTlA8*1 allele is a protective factor and UGTlA8*3 allele is a risk factor.