NF-kappa B expression in resected specimen of colonic cancer is higher compared to its expression in inflammatory bowel diseases and polyps.

NF-Kappa B has a significant role in inflammatory processes as well as in colorectal cancer. The aim of this study was to compare the expression of NF-kappa B in colonic adenocarcinoma specimen, colonic adenomas and inflammatory colonic tissues. Patients with colorectal cancer (CRC), colonic adenomas and inflammatory processes undergoing surgery were recruited. Following a routine pathological evaluation tissue samples were stained using anti NF-κB monoclonal antibodies. Expression of NF-κB was quantified using IMAGEJ program for immunohistochemistry staining. Samples were also stained and quantified for CEA expression. Fifty-six patients were included. 30 cancers, 6 polyps and 20 inflammatory processes. Expression of NF-κB was similar between polypoid and inflammation etiologies. However, it was significantly higher in CRC compared to both (p < 0.05). In cancer patients, NF-κB expression in the resection margins was correlated with positive node status. CEA expression was higher in the cancer group, less in the IBD group and the lowest in the colonic non diseased margins. Our results provide a supportive evidence that NF-κB pathway is strongly involved in colon cancer development and metastasis. Interestingly, expression of NF-κB in benign polypoid lesions was as high as in inflammatory etiologies. This support the role of NF-κB early in the adenoma to carcinoma sequence. Further research is needed to evaluate the exact role of NF-κB in tumor progression in order to look for diagnostic and therapeutic possibilities.

Phospholipase D activation mediates growth and migration of colon cancer cells interacting with cancer-associated fibroblasts.

Cancer-associated fibroblasts of the stroma play a major role in tumor promoting processes. In this study we evaluated the significance of Phospholipase D (PLD) enzyme activity in promoting human colon cancer malignant potency when interacting with proximal colonic fibroblasts. Human colon cancer cell lines SW480 and HCT116, and colonic fibroblasts CCD-18Co were used as an in vitro model. PLD's activity was measured in resting cancer cells and after culturing with fibroblasts and cancer-associated fibroblasts (CAFs) conditioned medium. The viability and migration level of the cancer cells alone and after co-culturing with fibroblast or CAFs conditioned medium were evaluated, with and without adding a PLD inhibitor. Exposure of colon cancer cells to CAFs conditioned medium significantly increased the level of PLD activity in the cancer cells (p<0.0001). Exposure of colon cancer to resting and activated fibroblast conditioned medium significantly enhanced the number of viable cancer cells as well as its migration level measured following 24 and 48 hours. Adding a PLD inhibitor significantly reduced the elevation of cell viability and migration of the colon cancer cells exposed to fibroblasts conditioned medium (p<0.005). In this in vitro model, inhibition of PLD significantly decreased proliferation and migration levels of colon cancer cells generated by stromal fibroblasts. This provides evidence that the PLD signaling pathway is directly involved in stroma-cancer interactions in the colon, thereby promoting cancer progression. Further research is needed in order to evaluate PLD as a target in colon cancer prevention or therapy.

Inhibition of TNFα in peritoneal fluids of patients following colorectal resection attenuates the postoperative stress-related increase in colon cancer cell migration: A prospective, in vitro study.

BACKGROUND AND OBJECTIVES: Surgical trauma in patients undergoing colorectal cancer resection generates local and systemic inflammatory responses that can affect oncological outcomes. Post-operative peritoneal fluids of patients undergoing colorectal surgery increase the pro-malignant effect of cancer cells in vitro with correlation to elevated TNFα in these fluids. This study evaluated whether inhibiting TNFα in patients' postoperative fluid biopsies would attenuate this effect. METHODS: Peritoneal fluids from 53 patients undergoing colorectal surgery were sampled before and daily after surgery via intra-abdominal drains. Fluid biopsies were evaluated for their impact on the migration capacity of colon cancer cells and for cytokine levels. TNFα was inhibited using infliximab and cell migration was reevaluated. RESULTS: Colon cancer migration capacity was increased in postoperative fluid biopsies from all patients (P < 0.005) and was elevated compared to pre-resection levels. Infliximab attenuated this effect in >90%, decreasing migration capacity by 30% (p < 0.001). CONCLUSIONS: Inhibition of TNFα in postoperative peritoneal fluids attenuates the increase in cancer cell migration capacity generated following colorectal resection. These findings correlate with other studies suggesting that attenuation of the post-operative inflammatory response may have oncological benefit. Clinical studies are needed to evaluate the effect of peri-operative TNFα inhibition in clinical settings.

The effect of bariatric surgery on hypothyroidism: Sleeve gastrectomy versus gastric bypass.

BACKGROUND: Hypothyroidism is prevalent in morbidly obese patients and may improve after a weight reduction surgery. OBJECTIVES: Laboratory and clinical changes in hypothyroid patients undergoing laparoscopic sleeve gastrectomy (LSG) or laparoscopic Roux-en-Y gastric bypass (LRYGB) were compared and evaluated. SETTINGS: Data were retrieved from a prospectively collected database of 2 public bariatric units. METHODS: Patients with hypothyroidism prior to bariatric procedure were evaluated for changes in thyroid stimulating hormone (TSH) and changes or cessation of hormone replacement therapy after surgery. Correlation between changes in TSH levels and percentage of excess weight loss and comparison between effects of LSG and LRYGB were evaluated. RESULTS: Ninety patients were included. Mean follow-up was 11 ± 9 .73 months. Mean body mass index decreased from 43.8 to 33.2 kg/m2. Forty patients had deranged elevated TSH levels prior to surgery that decreased significantly after surgery (mean 6.6 ± 1.9 to 2.9 ± 1.5 mU/L, P < .01). Of patients receiving hormone replacement therapy prior to surgery, 42% required lower doses, with a 61% mean decrease in doses, while 10% stopped hormone replacement therapy completely. No correlation was found between the improvement in TSH and percentage of excess weight loss. A significant advantage to one of the bariatric procedures (LSG [61] and LRYGB [29]) could not be established. CONCLUSIONS: LSG and LRYGB both proved to improve thyroid function in hypothyroid obese patients. No procedure was found to be superior. No correlation was found between percentage of excess weight loss and TSH reduction. This implies that the effect of bariatric surgery on the improvement of thyroid functions is mediated by mechanisms other than weight loss, probably hormonal.

Significant elevation of carcinoembryonic antigen levels in abdominal drains after colorectal surgery may indicate early anastomotic dehiscence.

BACKGROUND: Anastomotic leak after colorectal surgery is a severe complication leading to major postoperative morbidity and mortality. Leaks typically present on the 5th to 6th postoperative days; however, early anastomotic dehiscence occurs occasionally. This study evaluated carcinoembryonic antigen (CEA) levels in abdominal drains after colorectal resection to assess its potential as an early marker to predict anastomotic leaks. METHODS: This prospective study included 105 patients undergoing elective colorectal surgery. Fluids from the patients' abdominal drain system were sampled daily for up to 3 days after surgery and evaluated for CEA levels. RESULTS: Early anastomotic dehiscence occurred in 3 patients (2.8%) and was associated with a significant elevation of CEA in drain fluids (above 1,000 ng/mL). However, in patients who developed late leaks (7 patients, 6.7%) no significant elevation of CEA was observed. CONCLUSIONS: Significant elevation of CEA levels in abdominal drains in the early postoperative period may indicate early anastomotic dehiscence.

Postcolectomy Peritoneal Environment Increases Colon Cancer Cell Migration Capacity.

Background. Clinical data and animal models support an association between postoperative inflammatory response and the risk of colorectal cancer recurrence. Our aim was to evaluate postoperative peritoneal inflammation and its impact on cultured colon cancer cells' migration capacity. Methods. 23 patients undergoing elective colorectal resection with uneventful recovery were prospectively enrolled. Patients were operated on for both malignant and benign etiologies. Peritoneal fluids collected at surgery initiation and after surgery were evaluated for their effect on migration potential of human colon cancer cells using an in vitro scratch assay and on TNF-α, IL-1ß, IL-6, and IL-10 levels using bead-based fluorokine-linked multianalyte profiling. Results. Postoperative peritoneal fluid from all patients increased the migration capacity of colon cancer cells compared to preoperative levels. This effect was significant during the first two postoperative days and decreased thereafter. The increase in colon cancer cell migration capacity correlated with increased levels of peritoneal TNF-α and IL-10. Conclusion. In this pilot study, we have demonstrated that the intraperitoneal environment following colorectal resection significantly enhances colon cancer cells migration capacity. This effect is associated with postoperative intra-abdominal cytokines level. A larger scale study in colorectal cancer patients is needed in order to correlate these findings with perioperative parameters and clinical outcome.

Effect of sleeve gastrectomy on thyroid hormone levels.

BACKGROUND: Weight loss in morbidly obese patients is associated with changes in thyroid function. Studies have demonstrated equivalent changes following bariatric surgery. Changes in thyroid function were reported following laparoscopic Roux-en-Y gastric bypass (LRYGB), biliopancreatic diversion (BPD), and laparoscopic adjustable gastric banding (LAGB). No data exists on changes in thyroid function following laparoscopic sleeve gastrectomy (LSG). The aim of the current study is to evaluate changes in thyroid function following LSG in patients with normal thyroid function. METHODS: Data were retrieved from a prospectively collected database of patients who underwent LSG for morbid obesity. Euthyroid patients were evaluated for changes in TSH and free thyroxine (FT4), 6-12 months after surgery. Correlation between changes in thyroid hormone levels, excess weight loss (EWL), and baseline TSH were evaluated. RESULTS: Thirty-eight patients were included in the study. Mean BMI decreased from 42.4 to 32.5 kg/m(2) (P < 0.0001). Mean TSH levels decreased from 2.45 ± 0.17 mU/L at baseline to 1.82 ± 0.18 mU/L (P < 0.0001), whereas mean FT4 levels remained the same after surgery (13.27 ± 0.45 pmol/L compared to 12.96 ± 0.42 pmol/L, P = NS). TSH decrease was directly related to baseline TSH but did not correlate with EWL. CONCLUSIONS: This is the first study to evaluate changes in thyroid hormone levels following LSG for morbid obesity. TSH decrease and steady levels of FT4 are expected following LSG. These findings are comparable to reported changes following LRYGB. TSH decrease was not associated with EWL. Further studies are required to elucidate the exact mechanism of this effect.

Evaluation of peritoneal CEA levels following colorectal cancer surgery.

BACKGROUND: Peritoneal carcinoembryonic antigen (pCEA) levels in the early postoperative period following a curative resection of colorectal cancer (CRC) have not been previously studied. METHODS: Postoperative peritoneal fluids of 36 CRC patients followed by 24 benign colonic disease patients were evaluated for CEA levels and tumor cell presence. Serum CEA levels were also evaluated prior and after surgery. RESULTS: Although high postoperative pCEA levels were observed in some benign patients, more CRC patients exhibited significant elevation of postoperative pCEA (>5 ng/ml) compared to benign patients (50% vs. 23%, P = 0.039). Postoperative median pCEA levels of CRC patients were significantly higher compared to benign patients (5.4 vs. 2 ng/ml, P = 0.011). Specifically, pCEA levels in CRC patients were significantly elevated when measured during the first 24 hr after surgery. Postoperative pCEA levels were associated with colon tumor location compared to rectal location. However, no correlation was found with known risk factors for cancer recurrence or with serum CEA levels. CONCLUSIONS: Postoperative pCEA levels may be significantly elevated following a curative resection for CRC. Its significance within patient's prognostic evaluation remains to be studied. Inclusion of patient's follow-up data may reveal the significance of elevated pCEA levels following CRC resection.

Curcumin induces apoptosis and inhibits growth of orthotopic human non-small cell lung cancer xenografts.

Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related mortality. Curcumin is involved in various biological pathways leading to inhibition of NSCLC growth. The purpose of this study was to evaluate the effect of curcumin on expression of nuclear factor κB-related proteins in vitro and in vivo and on growth and metastasis in an intralung tumor mouse model. H1975 NSCLC cells were treated with curcumin (0-50 µM) alone, or combined with gemcitabine or cisplatin. The effects of curcumin were evaluated in cell cultures and in vivo, using ectopic and orthotopic lung tumor mouse models. Twenty mice were randomly selected into two equal groups, one that received AIN-076 control diet and one that received the same food but with the addition of 0.6% curcumin 14 days prior to cell implantation and until the end of the experiment. To generate orthotopic tumor, lung cancer cells in Matrigel were injected percutaneously into the left lung of CD-1 nude mice. Western blot analysis showed that the expressions of IkB, nuclear p65, cyclooxygenase 2 (COX-2) and p-ERK1/2 were down-regulated by curcumin in vitro. Curcumin potentiated the gemcitabine- or cisplatin-mediated antitumor effects. Curcumin reduced COX-2 expression in subcutaneous tumors in vivo and caused a 36% decrease in weight of intralung tumors (P=.048) accompanied by a significant survival rate increase (hazard ratio=2.728, P=.036). Curcumin inhibition of COX-2, p65 expression and ERK1/2 activity in NSCLC cells was associated with decreased survival and increased induction of apoptosis. Curcumin significantly reduced tumor growth of orthotopic human NSCLC xenografts and increased survival of treated athymic mice. To evaluate the role of curcumin in chemoprevention and treatment of NSCLC, further clinical trials are required.

Moringa Oleifera aqueous leaf extract down-regulates nuclear factor-kappaB and increases cytotoxic effect of chemotherapy in pancreatic cancer cells.

BACKGROUND: Fewer than 6% patients with adenocarcinoma of the pancreas live up to five years after diagnosis. Chemotherapy is currently the standard treatment, however, these tumors often develop drug resistance over time. Agents for increasing the cytotoxic effects of chemotherapy or reducing the cancer cells' chemo-resistance to the drugs are required to improve treatment outcome. Nuclear factor kappa B (NF-kB), a pro-inflammatory transcription factor, reportedly plays a significant role in the resistance of pancreatic cancer cells to apoptosis-based chemotherapy. This study investigated the effect of aqueous Moringa Oleifera leaf extract on cultured human pancreatic cancer cells - Panc-1, p34, and COLO 357, and whether it can potentiates the effect of cisplatin chemotherapy on these cells. METHODS: The effect of Moringa Oleifera leaf extract alone and in combination with cisplatin on the survival of cultured human pancreatic cancer cells was evaluated by XTT-based colorimetric assay. The distribution of Panc-1 cells in the cell cycle following treatment with Moringa leaf extract was evaluated by flow cytometry, and evaluations of protein levels were via immunoblotting. Data of cell survival following combined treatments were analyzed with Calcusyn software. RESULTS: Moringa Oleifera leaf extract inhibited the growth of all pancreatic cell lines tested. This effect was significant in all cells following exposure to ≥0.75 mg/ml of the extract. Exposure of Panc-1 cells to Moringa leaf extract induced an elevation in the sub-G1 cell population of the cell-cycle, and reduced the expression of p65, p-IkBα and IkBα proteins in crude cell extracts. Lastly, Moringa Oleifera leaf extract synergistically enhanced the cytotoxic effect of cisplatin on Panc-1 cells. CONCLUSION: Moringa Oleifera leaf extract inhibits the growth of pancreatic cancer cells, the cells NF-κB signaling pathway, and increases the efficacy of chemotherapy in human pancreatic cancer cells.

Association between tumorigenic potential and the fate of cancer cells in a syngeneic melanoma model.

The self-renewal potential of a cancer cell can be estimated by using particular assays, which include xenotransplantation in immunocompromised animals or culturing in non-adherent serum-free stem-cells media (SCM). However, whether cells with self-renewal potential actually contribute to disease is unknown. Here we investigated the tumorigenic potential and fate of cancer cells in an in-vivo melanoma model. We examined cell lines which were derived from the same parental line: a non-metastatic cell line (K1735/16), a metastatic cell line (K1735/M4) and a cell line which was selected in non-adherent conditions (K1735/16S). All cell lines exhibited similar proliferation kinetics when grown on culture plates. K1735/16 cells grown in soft agar or in suspension non-adherent conditions failed to form colonies or spheroids, whereas the other cell lines showed prominent colonogenicity and spheroid formation capacity. By using sphere limiting dilution analysis (SLDA) in serum-free media, K1735/16S and K1735/M4 cells grown in suspension were capable of forming spheroids even in low frequencies of concentrations, as opposed to K1735/16 cells. The tumorigenic potential of the cell lines was determined in SCID mice using intra footpad injections. Palpable tumors were evident in all mice. In agreement with the in-vitro studies, the K1735/M4 cell line exhibited the highest growth kinetics, followed by the K1735/16S cell line, whereas the K1735/16 cell line had the lowest tumor growth potential (P<0.001). In contrast, when we repeated the experiments in syngeneic C3H/HeN mice, the K1735/16 cell line produced macroscopic tumors 30-100 days after injection, whereas K1735/M4 and K1735/16S derived tumors regressed spontaneously in 90-100% of mice. TUNEL analysis revealed significantly higher number of apoptotic cells in K1735/16S and K1735/M4 cell line-derived tumors compared to K1735/16 tumors (P<0.001). The models we have examined here raised the possibility, that cells with high-tumorigenic activity may be more immunogenic and hence are more susceptible to immune-regulation.

Inhibition of cancer growth and induction of apoptosis by BGP-13 and BGP-15, new calcipotriene-derived vitamin D3 analogs, in-vitro and in-vivo studies.

One of the most innovative approaches to the treatment of cancer entails the use of 1α,25-dihydroxyvitamin D3 (calcitriol) analogs to inhibit cancer cell growth. We demonstrate here that BGP-13, a new calcipotriene-based 1α,25-dihydroxyvitamin D3 analog that we synthesized in our laboratory, inhibits the growth of prostate cancer (LNCaP), breast cancer (MCF-7), and colon cancer (HT-29) cell lines. Moreover, we also show that BGP-13 causes cells both to accumulate in G0-G1 and to activate caspase-3 and undergo apoptosis. In addition, we observed elevated vitamin D receptor (VDR) mRNA and protein levels in both LNCaP and MCF-7 cells following the exposure of the two cell lines to BGP-13. Importantly, we found that both the new analog BGP-13 and also BGP-15, another calcipotriene-based analog we synthesized previously and about which we published recently, inhibit the growth of HT-29 tumor xenografts in nude mice.

[The role of medicinal herbs with anti-inflammatory properties in prevention and treatment of cancer].

The association between chronic inflammation and carcinogenesis, as well as neoplastic progression, has been researched and is well-established. Being a central coordinator of immune responses, nuclear factor-kappa B (NFkappaB) signaling plays a critical role in cancer development and progression. The activation of the NFkappaB signaling pathway is highly monitored under normal conditions, and is mainly known as a key pathway in activation of immune responses. Constitutively active NFkappaB has been identified in most tumor cell lines, as well as in a wide variety of tumor tissues derived from cancer patients. Such activation may also affect the cancer's response to therapy, making it less susceptive to radio and chemo treatment. Hence, NFkappaB has become a target for inhibition by chemotherapeutic agents. Traditionally, medicinal herbs have been used to prevent and treat a variety of diseases, including cancer. In this article, we review several natural, herbal-derived compounds shown to have anti-inflammatory and anticancer activities mediated, at least in part, by NFkappaB signaling inhibition. Compounds of this sort may potentially serve as clinically effective anticancer treatments.

Induction of apoptosis and inhibition of prostate and breast cancer growth by BGP-15, a new calcipotriene-derived vitamin D3 analog.

The role of vitamin D3 in cancer prevention and its potential as an anticancer therapeutic agent have been researched and are well established. However, the clinical use of the natural vitamin D3 metabolite, 1alpha,25-dihydroxyvitamin D3 [1,25(OH)2D3 or calcitriol] is limited by a possible cause of hypercalcemia and hypercalciuria. A new 24-chloro calcipotriene-based vitamin D3 analog (BGP-15) was synthesized and examined for antiproliferative activity in the androgen-dependent cell lines of prostate cancer (LNCaP) and breast cancer (MCF-7). The new analog led to significant decrease in cell viability in cultured LNCaP and MCF-7 cell lines compared with calcipotriene and 1,25(OH)2D3. We observed elevated vitamin D receptor protein levels in both LNCaP and MCF-7 cells, which were treated with 5 micromol/l of 1,25(OH)2D3, calcipotriene or BGP-15 for 20 h, indicating vitamin D receptor-binding ability. Treatments of LNCaP and MCF-7 cells with 5 micromol/l BGP-15 and calcipotriene for 20 h generated procaspase-3 cleavage and therefore, apoptosis. Interestingly, BGP-15, and to a lesser extent calcipotriene, but not 1,25(OH)2D3, activated caspase-3 in MCF-7 cells, a cell line that normally lacks this specific caspase (and procaspase). It is presumed that management of MCF-7 with BGP-15 modulates procaspase-3 expression and cleavage, and a subsequent activation of caspase-3. Similar treatments of LNCaP cells induced procaspase-9 cleavage and therefore caspase-9 activation, whereas similar treatments of MCF-7 cells failed to induce caspase-9 activation. Cytochrome c release was, however, detected in both cell lines, LNCaP and MCF-7. In-vivo results suggested that BGP-15 (similar to its parent drug) did not cause calcium-related toxic side effects after chronic treatment.