Metastasis Of Colorectal Carcinoma Research Articles

Resection of hepatic metastatics from colorectal carcinoma. Factors related with recurrence and disease-free survival

Background: Surgery has become the standard treatment for patients with stage IV colorectal carcinoma and liver metastasis. Methods: From 2010-2015: 158 patients with liver metastases (142 CRC metastases that). We analyzed long-term results, actuarial survival and DFS, and compared risk factors with log-rank and chi-square (significance level p <0.05) Results: 72 metachronous (51%) and 70 synchronous metastases: 14 simultaneous resection, 14 liver-first, and colon-liver resection in 60%. With a mean follow-up of 23.7±16 months presented tumor recurrence 53(46.5%). At end, 55 patients were alive without tumor recurrence (41%). DFS at 1, 3 and 5 years is 65%, 37.9% and 32.1%, respectively, with an mean DFS interval of 33.6±2.96 months. Variables related to the recurrence: bilobar or segment I location,inverse surgery, second hepatectomies, presence of vascular invasion and R1 resection. Variables related to DFS: multicentric tumors, reverse surgery, rehepatectomies, vascular invasion and R1 resection. The use of preoperative chemotherapy, the type of resection or the need for transfusion has not been linked to long-term results. Conclusion: Resection of CCR hepatic metastases is the treatment of choice with 5-year DFS about 32% and mean DFS of 34 months. Initial hepatic resection in synchronous metastases has been associated with worse DFS as well as second resections. Neither the type of resection (major hepatectomy or atypical resection) nor the use or not of preoperative chemotherapy have been related to recurrence

A Case of Preoperatively Diagnosed Extrahepatic Bile Duct Metastasis of Colorectal Carcinoma

A Case of Preoperatively Diagnosed Extrahepatic Bile Duct Metastasis of Colorectal Carcinoma

Novel selective TOPK inhibitor SKLB-C05 inhibits colorectal carcinoma growth and metastasis

The mitogen-activated protein kinase (MAPK) signaling pathway member T-LAK cell–originated protein kinase/PDZ-binding kinase (TOPK/PBK) is closely involved in tumorigenesis and progression. Its overexpression in colorectal carcinoma (CRC) exacerbates tumor malignancy, promotes metastasis and results in dismal prognosis. Therefore, targeting TOPK is a promising approach for CRC therapy. Here, we report the development of a TOPK selective inhibitor SKLB-C05, with subnanomolar inhibitory potency. In vitro, SKLB-C05 exhibited excellent cytotoxicity and anti-migration and invasion activity on TOPK high-expressing CRC cells and induced cell apoptosis. These activities could attribute to its inhibition of TOPK downstream signaling including extracellular signal-regulated kinase 1/2 (ERK1/2), p38, and c-Jun N-terminal kinase 1, 2, and 3 (JNK1/2/3), as well as downregulation of FAK/Src- MMP signaling. Furthermore, SKLB-C05 disrupted cell mitosis and blocked CRC cell cycle. In vivo, oral administration of SKLB-C05 at concentrations of 20 and 10 mg kg−1·day−1 dramatically attenuated CRC tumor xenograft growth and completely suppressed hepatic metastasis of HCT116 cells, respectively. Thus, these findings suggest that SKLB-C05 is a specific TOPK inhibitor with potent anti-CRC oncogenic activity in vitro and in vivo.

Long noncoding RNA CRCMSL suppresses tumor invasive and metastasis in colorectal carcinoma through nucleocytoplasmic shuttling of HMGB2.

Long noncoding RNAs (lncRNAs) are pervasive transcripts that play pivotal roles in regulating chromatin dynamics, gene and protein expression. Aberrant expression and mutations of lncRNAs represent a driving force behind tumor invasion and metastasis, making them attractive cancer targets. However, most of the lncRNAs are still being discovered and conclusive experimental evidence for their functional relevance is still lacking for most malignancies. In this study, a differentially expressed lncRNA, designated as lnc-CRCMSL, is identified by microarray-based screenings on non-metastatic and metastatic CRC specimens. Lnc-CRCMSL is verified as an anti-metastatic gene and negatively correlated with the poor prognosis of CRC patients. Lnc-CRCMSL overexpression restricts tumor growth and metastasis in vivo and in vitro. Instead, lnc-CRCMSL silencing accelerates CRC cell proliferation and migration. RNA-pulldown assay identifies high mobility group box 2 (HMGB2) as a downstream protein of lnc-CRCMSL. Mechanically, lnc-CRCMSL physically binds to HMGB2 and stabilizes the localization of HMGB2 in the cytoplasm. Notably, lnc-CRCMSL knockdown lead to the shift of HMGB2 into nuclear, in which it triggers epithelial to mesenchymal transition (EMT) programming. Importantly, lnc-CRCMSL controls the cytoplasmic retention of HMGB2 and attenuates the interaction between HMGB2 and OCT4 to suppress EMT. Treatment of leptomycin B (LMB), a potent and specific nuclear export inhibitor, counteracts lnc-CRCMSL-mediated suppression of aggressive phenotypes and EMT process by accumulating the nuclear HMGB2. CONCLUSION: Our data highlight the anti-metastatic role of lnc-CRCMSL in stabilizing HMGB2 through lncRNA-protein interactions in the cytoplasm, and suggest that targeting lnc-CRCMSL may represent a therapeutic opportunity for managing metastatic CRC.

Antisense Oligonucleotides against miR-21 Inhibit the Growth and Metastasis of Colorectal Carcinoma via the DUSP8 Pathway

Accumulating research has documented that microRNA-21 (miR-21) plays an important role in the development of human colorectal carcinoma (CRC). Our recent work also showed that antisense oligonucleotides (ASOs) against miR-21 can impair the growth of CRC cells in vitro. However, the potential role of miR-21 in gene therapy against CRC remains to be fully elucidated. Here, we further observed the effect of ASOs against miR-21 on the growth and metastasis of CRC in vivo using a xenograft model of human CRC. We found that ASOs could effectively inhibit the growth and metastasis of CRC in vivo, accompanied by downregulated expression of miR-21 and reduced transduction of the AKT and ERK pathway. Mechanically, global gene expression analysis showed that the expression of DUSP8, a novel target of miR-21, was upregulated in tumor mass. Furthermore, overexpression of DUSP8 could remarkably suppress the proliferation and migration of CRC cells in vitro. Finally, downregulation of DUSP8 could abrogate the effects of ASOs against miR-21 on the proliferation and migration of CRC cells, as well as altered transduction of the AKT and ERK signaling pathway. Together, these data suggest that ASOs against miRNAs are an attractive and potential therapeutic for the treatment of human CRC and warrant further development.

The Prognostic Value of Cancer Stem Cell Markers (Notch1, ALDH1, and CD44) in Primary Colorectal Carcinoma.

Cancer stem cells proved to have a vital role in cell migration, invasion, metastasis, and treatment resistance of colorectal cancer (CRC) that subsequently lead to poor clinical outcomes. These stem cells may be a novel therapeutic target for the management of CRC progression. Signals of the Notch-1 pathway are responsible for acquisition of stem cell characters. ALDH1 and CD44 are usually detected in stem cells in colorectal cancer. The aims of this work are to evaluate the immunohistochemical expression of cancer stem cell markers ALDH1, Notch1, and CD44 in colorectal cancer and investigate their correlation with clinicopathological characters and patient survival. Paraffin-embedded specimens of 70 patients with primary colorectal carcinoma were analyzed for Notch 1, ALDH1, and CD44 expressions by immunohistochemistry. Notch1 was mainly located in the cytoplasm of CRC tissues, rarely expressed in adjacent normal tissues. A highly statistically significant relationship was found between grading, lymphovascular invasion, the degree of lymphocytic infiltration, peritumoral budding, lymph node ratio, lymph node metastasis, and Notch1 expression (p < 0.001). There was a highly statistically significant relationship found between AJCC stage and Notch1 expression (p < 0.001). CD44 was mainly located in the cell membrane of CRC tissues. A highly statistically significant relationship was found between grading (p = 0.006), lymphovascular invasion, the degree of lymphocytic infiltration, peritumoral budding, lymph node metastasis, lymph node ratio, and CD44 expression (p < 0.001). There was a highly statistically significant relationship found between AJCC stage and CD44 expression (p < 0.001). ALDH1 was detected in the cytoplasm of the CRC tissue. A highly statistically significant relationship was found between grading, lymphovascular invasion, the degree of lymphocytic infiltration, peritumoral budding, lymph node metastasis, lymph node ratio, and ALDH1 expression (p < 0.001). There was a highly statistically significant relationship found between AJCC stage and ALDH1 expression (p < 0.001). There is a highly statistically significant direct correlation between Notch1, CD44 expression, and ALDH1 expression (p < 0.001). There is a substantial correlation between Notch 1, ALDH1, and CD44 as cancer stem cell markers and lymph node metastasis, advanced stage and tumor recurrence in colorectal carcinoma. Expression of stem cell markers ALDH1, Notch1, and CD44 correlates with poor prognosis in a CRC and represents an independent prognostic factor. They are associated with a feature of epithelial-mesenchymal transition evidenced by their association with high tumor burden.

Hsp27 regulates epithelial mesenchymal transition, metastasis and proliferation in colorectal carcinoma.

The primary factor associated with poor survival rate in patients with colorectal carcinoma (CRC) is the presence of metastasis. The underlying molecular mechanisms of CRC metastasis are yet to be fully elucidated. The present study investigated the function of heat shock protein 27 (Hsp27) on the invasion and proliferation of CRC cells. The clinical significance of Hsp27 was evaluated using tissue microarray analysis (n=81). Invasion and metastasis assays were used to determine the function of Hsp27 in CRC metastasis in vitro and in vivo using RNA interference and the ectopic expression of Hsp27. The upregulation of Hsp27 has been frequently identified in CRC tissues. Patients with CRC and a high expression level of Hsp27 have a reduced overall survival rate. Silencing Hsp27 inhibited the growth and invasion of CRC cells in vitro and in vivo, whereas ectopic overexpression of Hsp27 promoted the proliferation and invasion of CRC cells in vitro. Furthermore, depletion of Hsp27 expression inhibited the epithelial-to-mesenchymal transition (EMT), whilst ectopic overexpression of Hsp27 induced EMT. The results of the present study indicated that Hsp27 serves an important function in the aggressiveness of CRC through inducing EMT. Hsp27 suppression may represent a potential therapeutic option for the suppression of CRC progression.

The Case Files

Figure: Image A.FigureFigureA 41-year-old woman presented to the emergency department with abdominal pain, noting multiple episodes over the past year of intermittent right upper quadrant pain that she described as “achy” and “fullness.” These episodes occurred sporadically with no identifiable exacerbating factors. Associated symptoms included a 20-pound unintentional weight loss over two months with occasional nausea and vomiting. She was a former smoker of 10-pack years and had stopped two months prior to arrival, and she reported drinking two to three glasses of wine a week. She did not have a primary care physician. Her vital signs on arrival were a temperature of 97.7°F, blood pressure of 132/97 mm Hg, heart rate of 98 bpm, 99% pulse ox on room air, respirations of 18 bpm, and weight of 76 kg. A physical exam revealed mild tenderness to deep palpation in the RUQ with appreciable hepatomegaly. Laboratory values were remarkable for a hemoglobin of 12.7 g/dL, mean corpuscular volume of 92, gamma-glutamyl transferase of 382 (9-48 U/L), total bilirubin of 0.6, alkaline phosphatase of 205 (44-147), aspartate aminotransferase of 55 U/L (10-37 U/L), alanine aminotransferase of 33 U/L (8-37 U/L). She was hemoccult-positive on rectal exam. Ultrasound showed multiple lobulated ill-defined masses throughout the liver. (Images A and B.) A CT abdomen/pelvis showed multiple heterogeneously enhancing irregular hepatic masses, with the largest measuring 10.6 x 10.1 x 11.1 cm. (Images C and D.) She was admitted for further evaluation. High Mortality Colorectal carcinoma is a common and lethal disease with around 140,000 cases diagnosed annually in the United States. (UpToDate, May 2, 2018; http://bit.ly/2IPlAS5.) About 56,000 patients die of the disease. Clinical research has shown that patients with a primary care physician have better management of chronic disease, access to preventive care, including cancer screenings, and chronic care of a multitude of medical problems. Advanced cases, like our patient's demonstrated, commonly present with iron-deficient anemia, rectal bleeding, abdominal pain, change in bowel habits, and sometimes an intestinal obstruction or perforation. A study of 388 patients diagnosed with colorectal carcinoma from 2011-2014 found the most common symptoms leading to a diagnostic colonoscopy to be rectal bleeding (37%), abdominal pain (34%), and anemia (23%). (UpToDate, May 2, 2018; http://bit.ly/2IPlAS5.) Clinical manifestations will differ depending on the location of the tumor. A change in bowel habits is more common in left-sided colorectal carcinoma, as is hematochezia. Iron-deficient anemia has been found to be more common in right-sided colorectal carcinoma. (UpToDate, May 2, 2018, http://bit.ly/2IPlAS5; Clin Colorectal Cancer 2016;15[1]:67; http://bit.ly/2KDxe7F.)Figure: Image B.Figure: Image C.The goal of laboratory evaluation is to assess patients' organ function for potential diagnostic and therapeutic procedures, as well as tumor burden. A baseline carcinoembryonic antigen carries a prognostic value. Patients with highly elevated values are associated with more advanced disease. (Anticancer Res 2001;21[4B]:3031.) Colorectal carcinoma metastases are most commonly found in the liver and the lung. Chemotherapy and radiation therapy following surgical resection of the primary tumor, when indicated, remain the mainstay of adjuvant therapy. Approximately 20 percent of patients are diagnosed with metastatic disease at their initial presentation. (UpToDate, May 2, 2018; http://bit.ly/2IPlAS5.) Our patient's colonoscopy showed an ulcerated sigmoid mass. The tumor marker carbohydrate Ag 19-9 was 670 U/mL (0-35). The confirmed diagnosis of metastatic adenocarcinoma with primary colorectal carcinoma was made based on subsequent CT-guided biopsy of the liver, followed by pathological report and a biopsy at the ulcerated sigmoid mass showing high-grade dysplasia. Given the extent and bilobar nature of the metastases, our patient's best option was to start systemic chemotherapy, followed by fully restaging after 12 cycles to determine if she would benefit from liver-directed therapy.Figure: Image D.

Heterogeneous expression of Lgr5 as a risk factor for focal invasion and distant metastasis of colorectal carcinoma.

Leucine-rich repeat-containing G-protein-coupled receptor 5 (Lgr5) is a downstream target gene of the Wnt/β-catenin signaling pathway and identified as a marker of cancer stem-like cells of colorectal carcinoma (CRC). Here, the heterogeneous expression pattern of Lgr5 and its clinical significance were studied by the method of immunohistochemistry in 204 CRC tumors at various pTNM stages. Lgr5 expression was found in 82.4% (168/204) cases, significantly more common in neoplastic cells at the infiltrative front (n = 59.5%, 110/185) or at the expanding front (n = 36.4%, 59/162) than at the tumor center (n = 16.7%, 34/204; P < 0.01). Tumor budding (TB) was discovered with significantly higher Lgr5 expression (n = 39.3%, 57/145, P = 0.03) and significantly positively correlated between Lgr5 expression and TB grade (r = 0.19, P = 0.02). Additionally, both positive Lgr5 expression and a high TB grade were significantly correlated to the depth of tumor invasion, lymph node metastasis, pTNM stage, and perineural invasion (P < 0.01). The study results suggest that heterogeneous expression of Lgr5 may be a risk factor for local invasion and distant metastasis of CRC.

Antiangiogenic Therapy in Colorectal Cancer.

Colorectal carcinoma is the third most common cancer worldwide. Approximately 20% of patients with colorectal cancer will have metastatic disease at the time of initial diagnosis, and approximately 30% to 50% of patients with primary colon cancer will relapse and die of metastatic cancer. The 5-year survival rate of metastatic colorectal cancer remains disappointing at approximately 10%.Angiogenesis plays a significant role in tumor growth and metastasis in colorectal carcinoma. There are currently 4 US Food and Drug Administration-approved antiangiogenic agents for metastatic colorectal cancer. Bevacizumab is the only antiangiogenic agent approved by the US Food and Drug Administration for first-line treatment of metastatic colorectal cancer. Other antiangiogenic agents include ramucirumab, ziv-aflibercept, and regorafenib. We review the data supporting the use of antiangiogenics in this disease.

Alcohol consumption promotes colorectal carcinoma metastasis via a CCL5-induced and AMPK-pathway-mediated activation of autophagy

There is a definite relationship between alcohol consumption and colorectal cancer (CRC) development. We investigated effect of alcohol consumption on CRC patients’ progression and prognosis by utilizing epidemiological data and found patients with alcohol consumption increased risks of tumor-node-metastasis (TNM), organ metastasis and poorer prognosis. Because their tumor tissues displayed increased expression of C-C chemokine ligand 5 (CCL5), we hypothesized CCL5 might participate in cancer progression in such patients. Ethanol increased the secretion of CCL5 in two CRC cell lines, HT29 and DLD-1. Treatment with CCL5 directly increased migratory ability of these cells, whereas neutralization or knockdown of CCL5 can partially block alcohol-stimulated migration. We further investigated underlying mechanism of CCL5-induced migration. Our results indicated that effects of CCL5 on migration are mediated by the ability of CCL5 to induce autophagy, a cellular process known to be critical for migration. Using high-throughput sequencing and western blotting, we found induction of autophagy by CCL5 takes place via AMPK pathway. Aforementioned ethanol increases CCL5 secretion, CCL5 activates autophagy through AMPK pathway, and autophagy increases migration was confirmed by experiments with autophagy or AMPK inhibitors. To sum up, our study demonstrates that chronic alcohol consumption may promote metastasis of CRC through CCL5-induced autophagy.

SATB2 is a supplementary immunohistochemical marker to CDX2 in the diagnosis of colorectal carcinoma metastasis in an unknown primary.

CDX2 is routinely used for identifying gastrointestinal origin of metastatic adenocarcinomas; but a high percentage of other carcinomas also show positivity with this antibody. SATB2 is a new immunohistochemical marker with a few studies showing that it is specifically expressed in a large majority of colorectal adenocarcinomas. We assessed SATB2 along with CDX2 in patient material with metastasis in order to determine whether the primary site could be identified as 'colon-rectum'. Metastasis in 67 liver biopsies, 108 lymph nodes from resection specimens and 36 serous effusions was analyzed retrospectively. Blinded slides stained for CDX2 and SATB2 were assessed individually by two pathologists and sensitivity, specificity and kappa statistics were calculated. Sensitivity for CDX2 in metastasis from colorectal adenocarcinomas was 93%; while in SATB2 it was 79%. The combination of CDX2 and SATB2 yielded a sensitivity of 79% and a high specificity of 93%. There was an acceptable level of agreement (κ=0.64) between the pathologists for both the markers in case of colorectal adenocarcinoma metastasis. CDX2 is a sensitive marker compared to SATB2; while the specificity of combination of CDX2 and SATB2 is high for metastasis from colorectal adenocarcinoma. SATB2 can be used as a supplementary marker along with CDX2 to identify colorectal origin for material received from patients clinically presenting with metastasis.

Abstract A17: Novel patient-derived xenograft (PDX) models from peritoneal metastasis of colorectal carcinoma for drug testing and biomarker analysis

Abstract Peritoneal metastasis (PM) represents a terminal tumor stage with limited therapy options for patients with colorectal cancer (CRC). To improve therapeutic efficacy and overall survival, availability of appropriate in vivo models of PM could improve the evaluation of chemosensitivity as well as identification of novel biomarkers and therapeutic targets to improve the therapeutic outcome of PM patients. In this context, we generated the first patient-derived xenograft (PDX) models of PM from CRC as platform to test chemotherapy response and to potentially analyze biomarkers. For the PDX establishment colorectal surgical specimens were subcutaneously (s.c.) transplanted onto immunocompromised NOG mice. The engrafted tumors were transferred to NMRI nu/nu mice for further passaging. They were characterized by histopathology, immunohistochemistry, and gene expression analyses using real-time RT-PCR. Chemosensitivity of PDX models was evaluated in vivo by application of a panel of conventional chemotherapeutic and of targeted drugs. For PDX establishment 68 CRC surgical specimens (taken from peritoneum and omentum) were transplanted, from which 22 PDX have engrafted. From those, 13 models of 10 patients have been analyzed. Their tumor doubling times ranged between 4 to 28 days. The histopathologic evaluation revealed maintenance of the original CRC histology in the PDX. The chemosensitivity testing of conventional and of targeted drugs in the 13 PM PDX models revealed the individual, diverging response of the PDX, such as for 5-FU, irinotecan, oxaliplatin, cetuximab, and erlotinib. More interestingly, different responses were observed in PDX of the omentum vs. peritoneum, which were derived from the same patient. Our results demonstrate that this novel panel of PDX maintains the morphology of the patient tumor in early passages, reflect heterogeneous response rates, and can be used as a preclinical in vivo platform for translational studies of potential clinical use. In conclusion, this PDX panel of PM of the CRC can be used for further studies to evaluate new biomarkers and to test novel therapies or combinations for PM of CRC. Citation Format: Wolfgang Walther, Eva Pachmayr, Bernadette Brzezicha, Britta Büttner, Beate Rau, Ulrike Stein. Novel patient-derived xenograft (PDX) models from peritoneal metastasis of colorectal carcinoma for drug testing and biomarker analysis [abstract]. In: Proceedings of the AACR Special Conference: Advances in Modeling Cancer in Mice: Technology, Biology, and Beyond; 2017 Sep 24-27; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(10 Suppl):Abstract nr A17.

Clinicopathological significance of G9A expression in colorectal carcinoma.

G9A, the primary histone methyltransferase (HMTase) for histone H3 lysine 9, is upregulated in numerous types of cancer and is critical for tumor cell proliferation. The present study aimed to investigate the G9A expression level in colorectal carcinoma (CRC) to evaluate the clinical significance of G9A in CRC. First, the present study detected the expression of G9A protein in 100 pairs of CRC specimens by immunohistochemistry staining and analyzed the correlations between G9A expression and pathological tumor features. It was found that G9A expression was increased markedly in CRC tumor specimens and the high expression was associated with tumor distant metastasis. Oncomine database analysis demonstrated an elevated expression level of G9A in various types of CRC. In total, 6 public available data sets from the Gene Expression Omnibus (GEO) were used and Gene set enrichment analysis (GSEA) was conducted. The results of the bioinformatics analysis demonstrated that high G9A expression was associated with American Joint Committee on Cancer staging, tumor differentiation, tumor relapse of CRC, and may serve a role in CRC cell proliferation. These findings suggested that G9A was overexpressed in CRC and involved in the tumorigenesis and distant metastasis of CRC. The expression level may also serve as a potential indicator for tumor recurrence in CRC. The present findings aided in the understanding of the crucial role of G9A in tumorigenesis and also offered novel ideas for CRC therapy.

Upregulation of c-mesenchymal epithelial transition expression and RAS mutations are associated with late lung metastasis and poor prognosis in colorectal carcinoma.

The present study aimed to investigate whether c-mesenchymal epithelial transition factor (C-MET) overexpression combined with RAS (including KRAS, NRAS and HRAS) or BRAF mutations were associated with late distant metastases and the prognosis of patients with colorectal cancer (CRC). A total of 374 patients with stage III CRC were classified into 4 groups based on RAS/BRAF and C-MET status for comprehensive analysis. Mutations in RAS/BRAF were determined using Sanger sequencing and C-MET expression was examined using immunohistochemistry. The associations between RAS/BRAF mutations in combination with C-MET overexpression and clinicopathological variables including survival were evaluated. In addition, their predictive value for late distant metastases were statistically analyzed via logistic regression and receiver operating characteristic analysis. Among 374 patients, mutations in KRAS, NRAS, HRAS, BRAF and C-MET overexpression were observed in 43.9, 2.4, 0.3, 5.9 and 71.9% of cases, respectively. Considering RAS/BRAF mutations and C-MET overexpression, vascular invasion (P=0.001), high carcino-embryonic antigen level (P=0.031) and late distant metastases (P<0.001) were more likely to occur in patients of group 4. Furthermore, survival analyses revealed RAS/BRAF mutations may have a more powerful impact on survival than C-MET overexpression, although they were both predictive factors for adverse prognosis. Further logistic regression suggested that RAS/BRAF mutations and C-MET overexpression may predict late distant metastases. In conclusion, RAS/BRAF mutations and C-MET overexpression may serve as predictive indicators for metastatic behavior and poor prognosis of CRC.

LIM and SH3 protein 1 knockdown suppresses proliferation and metastasis of colorectal carcinoma cells via inhibition of the mitogen-activated protein kinase signaling pathway.

LIM and SH3 protein 1 (Lasp-1), a focal adhesion protein, serves a critical role in the regulation of cell proliferation and migration. The role of Lasp-1, as well as the intracellular signaling pathways involved in metastasis and progression of colorectal carcinoma, remains unclear. In the present study, the regulatory effect of Lasp-1 and the underlying molecular mechanism involved in migration and invasion of colorectal carcinoma were investigated. RNA interference and overexpression in SW480 cells were performed to elucidate the role of Lasp-1. Reverse transcription-quantitative polymerase chain reaction, western blotting and immunofluorescence were conducted to determine the mRNA and protein levels of Lasp-1 and extracellular-signal-regulated kinase 1/2 (ERK1/2) in SW480 cells as well as tumor and adjacent normal tissues obtained from 20 patients with colorectal carcinoma. Furthermore, a cell proliferation assay, flow cytometric analysis, and cell migration and invasion assays were performed to examine the effect of Lasp-1 on cell growth. The results of the present study demonstrated that Lasp-1 and ERK1/2 were upregulated in tumor tissue compared with adjacent normal colorectal mucosa. Cell-based in vitro assays demonstrated that Lasp-1 knockdown suppressed the expression and activation of ERK1/2, whereas Lasp-1 overexpression resulted in ERK1/2 upregulation. Additionally, Lasp-1 knockdown inhibited cell proliferation, migration, and invasion and induced cellular apoptosis and G0/G1 cell-cycle arrest. The results of the present study indicate that Lasp-1 serves a critical role in the progression of colorectal carcinoma regulating the activation of the mitogen-activated protein kinase signaling pathway.

4:21 PM Abstract No. 331 Quantitative analysis of Lipiodol washout rates in intrahepatic cholangiocarcinoma and hepatic metastasis from neuroendocrine tumors and colorectal cancer after chemoembolization

To evaluate Lipiodol washout time after conventional transarterial chemoembolization (cTACE) in different hepatic malignancies including intrahepatic cholangiocarcinoma (ICC) as well as hepatic metastatic neuroendocrine tumors (NET) and colorectal carcinoma (CRC). This retrospective study included 41 patients, who underwent cTACE for the treatment of ICC (n = 9), NET (n = 14) or CRC metastases to the liver (n = 12). Lipiodol volume (LV) in cm3 was analyzed on follow-up non-contrast CT imaging obtained within 24 hours (LV1) as well as 40-220 days (LV2) after cTACE using a volumetric segmentation and image analysis software (qEASL; Medisys, Philips Research, Suresnes, France). The lipiodol washout rate was calculated as the ratio of (LV1-LV 2) to the interval (in days) between both CT scans. Additionally, tumor response on contrast-enhanced MRI was assessed 1 month after cTACE according to qEASL criteria. Lipiodol was visible on all follow-up CT scans. The LV2 was significantly lower than LV1 in all three tumors types (p<0.001). The mean lipiodol washout rate was -0.034 for ICC, -0.045 for NET, and -0.034 cm3/day for CRC metastases. No significant difference in lipiodol washout rates was observed between the three types of tumors (p>0.05). As for tumor response, 2/9 ICC patients, 4/14 NET patients and 5/18 CRC patients demonstrated response to cTACE according to qEASL criteria. However, no difference in lipiodol washout rate was revealed between responders and non-responders (p>0.05). This study showed that lipiodol washout rates after cTACE were similar for different hepatic malignancies including ICC, NET and CRC metastases. All observed tumors showed consistent washout regardless of response suggesting that lipiodol washout rate may not be predictive of tumor response to treatment.

Membranous or Cytoplasmic HER2 Expression in Colorectal Carcinoma: Evaluation of Prognostic Value Using Both IHC & BDISH

ABSTRACTBackground: Human epidermal growth factor recptor-2 (HER2) was identified as a driver gene in several types of cancers with both prognostic and predictive value. However, the molecular association of HER2 gene mutation with HER2 gene amplification and/or protein expression in cancer tissues has not been clearly defined. Moreover, there is little information available on HER2 status role in tumor progression and metastasis in colorectal carcinoma (CRC) compared to other solid tumors. The aim of this study was to evaluate both HER2 amplification and protein expression profiles using immunohistochemistry (IHC) and bright-field dual in situ hybridization (BDISH) techniques, respectively. Patients and methods: Tissue microarray (TMA) was constructed to accommodate a total of 243 CRC formalin-fixed paraffin embedded (FFPE) samples of consent patients and stained by IHC and BDISH methods. The expression patterns of HER2 protein status were evaluated and correlated to HER2 gene amplification status and then assessed for its prognostic value. Results: The expression profile of 58% samples showed cytoplasmic expression patterns of different categories. Interestingly, only 1% showed strong (+3) membranous expression pattern of HER2 with perfect match with their corresponding gene amplification status (>2). However, the cytoplasmic HER2 protein status did not show significant correlation with most clinicopathological features and survival outcomes except with age (p = 0.04) and tumor size (p = 0.03). Conclusion: We demonstrated that the membranous HER2 gene/protein status is infrequent, while the main fraction of HER2 overexpression was cytoplasmic and lacking prognostic value. This cytoplasmic HER2 overexpression was induced through a gene-amplification independent pathway, making the HER2 gene status evaluation approach in those cases not worthy. Further investigations about the molecular pathways of the cytoplasmic HER2 protein in CRC and its associations with survival outcomes are required to allow either a breakthrough in CRC management; or to confirm the hypothesis of a marginal role in CRC onset and progression.

Association of microRNA-423 rs6505162 C>A polymorphism with susceptibility and metastasis of colorectal carcinoma.

Studies have evaluated the association between the SNP miRNA-423 rs6505162 C>A and cancer risk in several cancers with contradictory outcomes. It was reported that miRNA-423 rs6505162 C>A polymorphism was associated with the overall survival and the recurrence-free survival of colorectal carcinoma. However, no studies have reported the association between miRNA-423 rs6505162 C>A polymorphism and susceptibility of colorectal carcinoma.In this study, we investigated the association between miRNA-423 polymorphism with risk and clinicopathological parameters of colorectal carcinoma. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to genotype 117 colorectal carcinoma patients and 84 healthy controls.Our data indicated the frequencies of rs6505162 genotypes and alleles were significantly different between colorectal carcinoma patients and controls. Compared with CC homozygote, the AC heterozygote exhibited a significantly decreased risk of colorectal carcinoma; and the combination of AC and AA genotype was associated with decreased risk of colorectal carcinoma. The allele distribution of rs6505162 was significantly different between cases and controls. Furthermore, miR-423 rs6505162 C>A genotype showed a significant association with metastasis in patients (P = .022).Our study suggested that miR-423 rs6505162 C>A polymorphism was associated with the susceptibility and metastasis of colorectal carcinoma, and that miR-423 rs6505162 C>A polymorphism might be a potential biomarker for colorectal carcinoma.

Predictive Factors for Lymph Node Metastasis in Submucosal Invasive Colorectal Carcinoma: A New Proposal of Depth of Invasion for Radical Surgery.

Patients with lymph node metastasis (LNM) in submucosal invasive colorectal carcinoma (SM cancer) require additional surgical treatment after endoscopic dissection. However, because additional radical resection after endoscopic local resection may be unnecessary for cases without LNM, more specific criteria are required in order to diminish the incidence of further radical resection after endoscopic dissection. A total of 492 patients with biopsy-proven SM cancer who underwent curative surgery between January 2008 and December 2012 were collected and were divided into LNM group and no LNM group. The cutoff value for the depth of submucosal invasion was analyzed by a receiver operating characteristic (ROC) curve. In this retrospective study, the association between LNM and clinicopathologic factors was analyzed by logistic regression analysis. The depth of submucosal invasion of 1900μm was determined as the cutoff value by ROC curve. Significant, independent predictive factors for LNM included the depth of submucosal invasion>1900μm (odds ratio [OR] 7.5; 95% confidence interval [CI] 3.1-18.3; p<0.001), venous invasion (OR 2.4; 95% CI 1.1-5.5; p=0.03), and poorly differentiated/mucinous adenocarcinoma (OR 6.3; 95% CI 1.3-30.8; p=0.02). Our study demonstrates that the depth of submucosal invasion (>1900μm), vascular invasion and poorly differentiated/mucinous carcinoma were predictive factors of LNM in patients with SM cancer. These predictors may help to reduce the incidence of unnecessary surgery after endoscopic resection.