Assessing the Therapeutic Impacts of HAMLET and FOLFOX on BRAF-Mutated Colorectal Cancer: A Study of Cancer Cell Survival and Mitochondrial Dynamics In Vitro and Ex Vivo.

Background and Objectives: Colorectal cancer (CRC) is a major global health challenge. The BRAF V600E mutation, found in 8-12% of CRC patients, exacerbates this by conferring poor prognosis and resistance to therapy. Our study focuses on the efficacy of the HAMLET complex, a molecular substance derived from human breast milk, on CRC cell lines and ex vivo biopsies harboring this mutation, given its previously observed selective toxicity to cancer cells. Materials and Methods: we explored the effects of combining HAMLET with the FOLFOX chemotherapy regimen on CRC cell lines and ex vivo models. Key assessments included cell viability, apoptosis/necrosis induction, and mitochondrial function, aiming to understand the mutation-specific resistance or other cellular response mechanisms. Results: HAMLET and FOLFOX alone decreased viability in CRC explants, irrespective of the BRAF mutation status. Notably, their combination yielded a marked decrease in viability, particularly in the BRAF wild-type samples, suggesting a synergistic effect. While HAMLET showed a modest inhibitory effect on mitochondrial respiration across both mutant and wild-type samples, the response varied depending on the mutation status. Significant differences emerged in the responses of the HT-29 and WiDr cell lines to HAMLET, with WiDr cells showing greater resistance, pointing to factors beyond genetic mutations influencing drug responses. A slight synergy between HAMLET and FOLFOX was observed in WiDr cells, independent of the BRAF mutation. The bioenergetic analysis highlighted differences in mitochondrial respiration between HT-29 and WiDr cells, suggesting that bioenergetic profiles could be key in determining cellular responses to HAMLET. Conclusions: We highlight the potential of HAMLET and FOLFOX as a combined therapeutic approach in BRAF wild-type CRC, significantly reducing cancer cell viability. The varied responses in CRC cell lines, especially regarding bioenergetic and mitochondrial factors, emphasize the need for a comprehensive approach considering both genetic and metabolic aspects in CRC treatment strategies.

Predictors of Quality of Life Six Years after Curative Colorectal Cancer Surgery: Results of the Prospective Multicenter Study.

Background and objectives: Improving early diagnosis and advances in colorectal cancer (CRC) treatment leads to longer survival of these patients. The purpose of this study was to identify the main surgical factors affecting long-term Quality of life (QoL) among colorectal cancer patients after surgery. Materials and Methods: QoL was prospectively evaluated in patients undergoing elective colorectal cancer resection operations in three CRC surgery centers of Lithuania using EORTC generic (QLQC-30) and disease-specific (QLQ-CR29) questionnaires at the time of preoperative admission and 1, 24, and 72 months after surgery. QoL was evaluated among different patient groups, diagnostic and treatment modalities, disease, and postoperative complications. Non-parametric tests and multivariate logistic regression models were used for statistical analysis. Results: Eighty-eight consecutive CRC patients from three institutions were included in the study over a three-month inclusion period, 42 (47.73%) women and 46 (52.27%) men, mean age 64.2 ± 11.5 years. Most tumors were localized in the sigmoid colon and rectum. The largest number of patients had stage III cancer. Twenty-nine patients died-a 6-year survival rate was 67%. 50 of 59 live patients (84.8%) responded to the questionnaire 6 years after their operation. Evaluating changes in quality of life 72 months after surgery with assessments before surgery, both questionnaire responses revealed good long-term CRC surgical treatment results: improved general and functional scale estimates and decreased symptom scale ratings. The multivariate analysis found that age, stoma formation, and rectal cancer were independent risk factors for having worse QoL six years after surgical intervention. Conclusions: Six years after surgery, QoL returns to preoperative levels. Age, stoma formation, adjuvant treatment, and rectal cancer reduce long-term QoL.

The Role of VEGFA, COX2, HUR and CUGBP2 in Predicting the Response to Neoadjuvant Therapy in Rectal Cancer Patients.

Background and objectives: The effectiveness of neoadjuvant therapy, which is commonly used for stage II-III rectal cancer (RC) treatment, is limited. Genes associated with the pathogenesis of RC could determine response to this treatment. Therefore, the aim of this study was to investigate the potential predictive value of VEGFA, COX2, HUR and CUGBP2 genes and the associations between post-treatment changes in gene expression and the efficacy of neoadjuvant therapy. Materials and Methods: Biopsies from RC and healthy rectal tissue of 28 RC patients were collected before neoadjuvant therapy and 6-8 weeks after neoadjuvant therapy. The expression levels of VEGFA, COX2, HUR, CUGBP2 genes were evaluated using a quantitative real-time polymerase chain reaction. Results: The results reveal a significantly higher expression of VEGFA, COX2 and HUR mRNA in RC tissue compared to healthy rectal tissue (p < 0.05), and elevated VEGFA gene expression in pre-treatment tissues was associated with a better response to neoadjuvant therapy based on T-stage downstaging (p < 0.05). The expression of VEGFA, HUR and CUGBP2 genes significantly decreased after neoadjuvant therapy (p < 0.05). Responders to treatment demonstrated a significantly stronger decrease of VEGFA and COX2 expression after neoadjuvant therapy than non-responders (p < 0.05). Conclusions: The findings of this study suggest that the pre-treatment VEGFA gene expression might have predictive value for the response to neoadjuvant therapy, while the post-treatment decrease in VEGFA and COX2 gene expression could indicate the effectiveness of neoadjuvant therapy in RC patients.

Improved perioperative care is associated with improved long-term survival in colorectal cancer.

PURPOSE: To compare perioperative colorectal cancer care and survival in patient cohorts operated in 2005 and in 2010 in Lithuania. METHODS: Comparative observational cohort study was performed. The study was conducted in the three Lithuanian cancer hospitals. Patients, who underwent curative surgery for colorectal cancer in 2005 and 2010, were included. Demographic characteristics, distribution of the tumors, preoperative diagnostics and staging, surgical treatment, the quality of pathological examination, morbidity, and mortality were analyzed. One- and 5-year overall survival data were compared between the groups. RESULTS: Colorectal cancer diagnostics and treatment improved from 2005 to 2010 significantly. The disease was identified as stage III-IV for 45 vs. 48% of the patients; however, computed tomography staging scan was performed only for 5.9 vs. 17.8% in 2005 and 2010, respectively. Laparoscopic operations were performed 1.5 vs. 10.5% and abdominoperineal resections-42.7 vs. 31.7% in 2005 and 2010, respectively. The number of harvested lymph nodes was mentioned in 55.8 vs. 97.7% of the cases, whereas more than 12 lymph nodes were examined in 18 vs. 66.6% of cases after histological examination. The overall 5-year survival was 52.1 vs. 63.1% (p < 0.0001), while the 5-year survival of the patients with stage IV of disease was 4.2 vs. 17.8% in 2005 and 2010, respectively. CONCLUSION: Preoperative investigation, surgical treatment, pathological examination, and postoperative course are associated with improved overall survival in colorectal cancer patients, undergoing curative surgery in the resource-limited settings.

Long-term Survival of Patient with Ampulla of Vater Metastasis of Renal Cell Carcinoma.

Ampulla of Vater metastases from renal cell carcinoma are rare. The time between detection of the primary tumour and its metastasis may extend to years. Management should be aggressive, since the prognosis of renal cell carcinoma is unpredictable and curative surgery of metastases may extend patient survival and even lead to definite cure. Herein we report a case of long-term survival after successful surgical treatment of a renal cell carcinoma metastasis to the ampulla of Vater. A 62-year-old man with a history of renal cell carcinoma in the left kidney underwent a successful left nephrectomy. Eight months later duodenoscopy showed a tumour at the site of papilla of Vater. Biopsy confirmed the diagnosis of carcinoma. Contrast enhanced computer tomography scan verified the periampullary mass, dilatation of the pancreatic and the common bile duct. No radiological signs of either local advancement or distant metastases were present. Pylorus-preserving pancreatoduodenectomy with lymphadenectomy was performed. Pathology report disclosed metastatic lesions in the papilla of Vater from the clear cell carcinoma of the kidney. The postoperative course was uneventful, and the patient lived for 14 years after pancreatoduodenectomy and, following thorough investigations, was free from local and systemic recurrence. Pancreatoduodenectomy can provide long-term survival in selected cases with solitary papilla of Vater metastasis from renal cell carcinoma. Favourable long-term survival rates suggest that these patients should be considered candidates for pancreatoduodenectomy if experienced pancreatic surgeon is available and no other metastases are found.

Preoperative conventional chemoradiotherapy versus short-course radiotherapy with delayed surgery for rectal cancer: results of a randomized controlled trial.

BACKGROUND: There still is no evidence which neoadjuvant therapy regimen for stage II-III rectal cancer is superior. The aim of this study was to compare results achieved after long-course chemoradiotherapy (CRT) with short-term radiotherapy (RT) followed by delayed surgery. METHODS: A randomized trial was carried out between 2007-2013. One hundred fifty patients diagnosed with stage II-III rectal cancer were randomized into one of two neoadjuvant treatment arms: conventional chemoradiotherapy (CRT) and short-term radiotherapy (RT) followed by surgery after 6-8 weeks. Primary endpoints of this trial were downstaging and pathological complete response rate. Secondary endpoints were local recurrence rate and overall survival. RESULTS: The pathological complete response was found in 3 (4.4%) cases after RT and 8 (11.1%) after CRT (P = 0.112). Downstaging (stage 0 and I) was observed in 21 (30.9%) cases in RT group vs. 27 (37.5%) cases in CRT group (P = 0.409). Median follow-up time was 39.7 (range 4.9-79.7) months. 3-years overall survival (OS) was 78% in RT group vs. 82.4% in CRT group (P = 0.145), while disease-free survival (DFS) differed significantly - 59% in RT group vs. 75.1% in CRT group (P = 0,022). Hazard ratio of cancer progression for RT patients was 1.93 (95% CI: 1.08-3.43) compared to CRT patients. CONCLUSION: Three-years disease-free survival was better in CRT group comparing with RT group with no difference in overall survival. TRIAL REGISTRATION: http://clinicaltrials.gov identifier NCT00597311. January 2008.

Short-term results of quality of life for curatively treated colorectal cancer patients in Lithuania.

BACKGROUND AND OBJECTIVE: Treatment options for colorectal cancer patients create the need to assess the quality of life (QoL) of colorectal cancer patients in the early postoperative period when changes are potentially greatest. The aim of the current study was to assess the QoL of colorectal cancer patients following open and laparoscopic colorectal surgery. MATERIALS AND METHODS: A total of 82 consecutive patients requiring elective open or laparoscopic colorectal surgery were recruited to the study for 3 months in the three colorectal surgery centers of Lithuania. Patients completed the EORTC QLQ-C30 (version 3.0) questionnaire before surgery, 2 and 5 days, 1 and 3 months after operation. The EORTC QLQ-CR29 questionnaire was completed before surgery and at 1 and 3 months after operation. Analysis was done according to the manual for each instrument. RESULTS: EORTC QLQ-C30 reflected the postoperative recovery of QoL. The global health status, cognitive and emotional functioning came back to the preoperative level in one month after operation. Physical and role functioning for laparoscopic group was significantly improved in 1 month after operation and in 3 months for open surgery group respectively. Colorectal module EORTC-QLQ-CR29 found that future perspective increased significantly in laparoscopic group 1 month after operation. CONCLUSIONS: The present study showed that majority of functional scale scores came back to the preoperative level during the first 3 months after colorectal cancer surgery. Differences in QoL according to surgical approach are mostly expressed on this period.

Challenges in diagnosing an extraosseous Ewing sarcoma: A case report.

INTRODUCTION: Ewing sarcoma is a highly malignant and rare tumour of bones and soft tissue. It may occur at any age, but it is more common in children and teenagers. CASE REPORT: We report a case of a 56-year-old woman with EES involving the right iliac fossa. Previous abdominal trauma with retroperitoneal hematoma, nonspecific symptoms and unusual age for EES have caused diagnostics difficulties. The first histopathological examination misdiagnosed tumour to be a GIST, and just after the second surgery the accurate diagnosis of EES was made. CONCLUSIONS: The diagnosis of ES sometimes is complicated and delayed. Prompt detailed examination and imaging studies should be performed to people with long lasting pain without trauma and other nonspecific symptoms, especially followed by a palpable mass. The treatment of EES is multimodal.

Lymph node retrieval after resection of rectal cancer following preoperative chemoradiotherapy.

BACKGROUND: According to the current guidelines of proper TNM staging, 12 lymph nodes per specimen are crucial. This study assessed the role of preoperative radiochemotherapy on the number of lymph nodes detected in the tumor-bearing specimen. MATERIAL AND METHODS: Retrospective data of 138 patients who underwent surgery for stage II and III rectal cancer without preoperative radiochemotherapy during the period of 2004-2006 (control group) were compared with prospective data of 38 patients who received preoperative radiochemotherapy during the period of 2007-2008 (study group). The number of patients with metastatic lymph nodes, number of lymph nodes per specimen, number of metastatic lymph nodes per specimen, and the size of the tumor between the groups were compared. RESULTS: Positive lymph nodes were detected in 88 (64%) patients in the control group as compared with 9 (21%) patients in the study group (P<0.05). The mean number of lymph nodes per specimen in the control group was 13.5, while in the study group, the mean number of lymph nodes per specimen was 6.29 (P<0.05). There was a significant difference in the mean number of metastatic lymph nodes per specimen between the groups (5.12 in the control group versus 2.11 in the study group; P<0.05). The mean size of the tumor was 4.37 cm in the control group and 2.45 cm in the study group (P<0.01). CONCLUSIONS: Preoperative radiochemotherapy for advanced rectal cancer significantly decreased the number of lymph nodes detected in the tumor-bearing specimen. This also resulted in a significant decrease in the number of metastatic lymph nodes detected in the specimen, and fewer patients with stage III (N+) cancer were diagnosed. Preoperative radiochemotherapy could induce a significant downsizing and downstaging of advanced rectal cancer, but great care in operative and pathologic examination techniques must be taken to ensure appropriate staging.

Percutaneous transhepatic biliary stenting: the first experience and results of the Hospital of Kaunas University of Medicine.

Malignant biliary obstruction may be caused by cholangiocarcinoma and other nonbiliary carcinomas. At the time of diagnosis, 90% of patients with malignant obstructive jaundice may benefit from palliative treatment only. The objective of palliation is to relieve jaundice-related symptoms, prevent cholangitis, prolong survival, and improve quality of life. Percutaneous transhepatic biliary stenting is a well-established procedure used in patients with malignant obstruction of intra- and extrahepatic bile ducts. Twelve patients (9 women, 3 men; mean age, 68 years; range, 44-88 years) with inoperable malignant biliary obstruction were selected for percutaneous transhepatic biliary stenting with metallic stents in the period from January to December 2007. Technical and clinical success rate in this patient series was 83% and 80%, respectively. Minor and major complications occurred in 17% and 8% of cases, respectively, which is in the range reported by the others. This is our first experience of percutaneous transhepatic biliary stenting at the Hospital of Kaunas University of Medicine and, to our knowledge, the first reported patient series in Lithuania. These first results encourage expanding effective palliation by the employment of the percutaneous transhepatic biliary stenting in patients with nonresectable malignant biliary obstruction or in case of a recurrent disease after curative surgery. The cost effectiveness of percutaneous transhepatic biliary stenting against percutaneous transhepatic biliary drainage has yet to be evaluated in a prospective manner. However, immediate clinical benefits and positive short-term outcomes are unequivocal.

Molecular fingerprinting and autocrine growth regulation of endothelial cells in a murine model of hepatocellular carcinoma.

In a mouse model of hepatocellular carcinogenesis, highly vascularized tumors develop through two distinct morphologic phases of neovascularization. We show that increased vascular caliber occurs first, followed by extensive vessel sprouting in late-stage carcinomas. To define molecular pathways in tumor neovascularization, endothelial cells were directly purified from normal liver and advanced tumors. Gene expression profiling experiments were then designed to identify genes enriched in the vascular compartment. We report that Cathepsin S is the major protease specifically overexpressed during vessel sprouting. We also show that the CC chemokines CCL2 and CCL3 are secreted by neovessels and stimulate proliferation through their cognate receptors in an autocrine fashion. This suggests that chemokine signaling represents the most prominent signaling pathway in tumor-associated endothelial cells and directly regulates vessel remodeling. Furthermore, high angiogenic activity is associated with attenuated lymphocyte extravasation and correlates with expression of the immunomodulatory cytokine interleukin 10. This is the first comprehensive study addressing liver-specific vascular changes in a murine autochthonous tumor model. These novel insights into liver angiogenesis infer an environmental control of neovascularization and have important implications for the design of antiangiogenic therapies.

Active leukocyte crawling in microvessels assessed by digital time-lapse intravital microscopy.

OBJECTIVE: The ability of active movement is an important feature of leukocytes. Here, we used a hybrid technique that combines intravital microscopy and digital time-lapse video microscopy to investigate the physiology and molecular mechanisms of intravascular leukocyte movement. METHODS: Intravital microscopy of mesenteric venules was performed in male, Wistar rats using digital video recording and time-lapse image compression. The leukocyte movement and extravasation were analyzed after local application of TNF-alpha, after blockade of endothelial (anti-ICAM-1 antibody) and leukocyte (anti-CD18 antibody) adhesion molecules. Additionally, the migratory activity of isolated leukocytes in collagen gel was analyzed and compared with their intravascular locomotion. RESULTS: Adherent leukocytes showed an active intraluminal crawling along the endothelial lining. Most permanent stickers (84 +/- 13%) crawled actively on the intraluminal site of venules. Baseline measurement of leukocyte crawling velocity yielded an average 9.0 +/- 1.8 mum/min that was not significantly different from crawling velocity of extravascular leukocytes (8.9 +/- 4.5 mum/min). The maximum distance of leukocyte crawling observed was 150 microm. The maximum time of crawling was 15 min. Intraluminal crawlers traveled over a mean distance of 35 +/- 17 mum with the average duration of 5.4 +/- 1.4 min. Under unstimulated conditions, almost all crawling leukocytes detached from the endothelium and did not migrate through the vascular wall. TNF-alpha induced a significant increase of leukocyte extravasation. Anti-ICAM-1 and anti-CD18 antibodies significantly reduced leukocyte crawling. The proportion of isolated migrating leukocytes in collagen gel (87% +/- 6%) was not significantly different from the percentage of intravascular crawling leukocytes in vivo. CONCLUSIONS: The method of digital time-lapse intravital microscopy represents an advantageous technology for the investigation of intravascular, transendothelial, and extravascular migration of leukocytes. Using this technology, we showed that leukocyte-endothelial-interactions are an active and dynamic process. This process involves long-time (several minutes) crawling of leukocytes along the endothelium and, finally, detachment from the endothelium. Intravascular leukocyte crawling reflects the migratory potential of circulating leukocytes and strongly depends on the expression of adhesion molecules. For extravasation, an additional pro-inflammatory stimulus is required.