Irinotecan plus raltitrexed as second-line treatment in locally advanced or metastatic colorectal cancer patients: a prospective open-label, single-arm, multi-center, phase II study.

BACKGROUND: Colorectal cancer is the third most common cancer and the second leading cause of cancer death. There are limited therapeutic options for the treatment of locally advanced or metastatic colorectal cancers which fail first-line chemotherapy. Phase I/II studies showed that the combined application of the raltitrexed and irinotecan has significant synergistic effect and acceptable toxicity. However, most of these previous studies have relatively small sample size. METHODS: This is a prospective open-label, single-arm, multi-center, Phase II trial. Brief inclusion criteria: patients were aged 18 to 75 years with locally advanced or metastatic colorectal cancer after failure of 5-FU and oxaliplatin therapy. Enrolled patients received raltitrexed (3 mg/m2, d1) and irinotecan (180 mg/m2, d1) each 21-day cycle until disease progression or unacceptable toxicity. The primary endpoint was progression-free survival, and the secondary endpoints were disease control rate, objective response rate, overall survival and safety. RESULTS: A total of 108 patients were enrolled between September 2016 and May 2020. The median age was 61 years, ECOG 1 score accounts for 67.6%, the rest were ECOG 0. A total of 502 cycles were completed, with an average of 4.6 cycles and a median of 4 cycles. 108 patients were evaluated, with an objective response rate of 17.6%, and disease control rate of 76.9%. The median follow-up time was 27 months (range:3.1-61.0 m) at data cut-off on March 2023. Median progression-free survival was 4.9 months (95% CI 4.1-5.7) and median overall survival was 13.1 months (95% CI 12.2-15.5). The most common adverse events that were elevated are alanine aminotransferase increased, aspartate aminotransferase increased, fatigue, diarrhoea, neutrocytopenia, thrombocytopenia, hypohemoglobin, and leukocytopenia. Most of the adverse events were Grade I/II, which were relieved after symptomatic treatment, and there were no treatment-related cardiotoxicities and deaths. CONCLUSIONS: The combination of raltitrexed and irinotecan as second-line treatment for mCRC could be a reliable option after failure of standard 5-Fu-first-line chemotherapy in locally advanced or metastatic colorectal cancers, especially for patients with 5-FU intolerance (cardiac events or DPD deficiency patients). TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT03053167, registration date was 14/2/2017.

Case report: A case of rare metastasis of gastric cancer to the axillary lymph node metastasis treated with combination immunotherapy.

Lymph node (LN) metastasis is a common mode of metastasis in advanced gastric cancer (GC), while axillary LN metastasis infrequently occurs in GC. There are few reports on this rare type of metastasis - especially its clinicopathological features - and systemic treatment are unclear. We describe a case of GC with extensive metastasis, including the rare axillary LN metastasis. The patient achieved partial response of optimal efficacy, who was treated with combination immunotherapy as second-line treatment for nearly two years. The potential mechanisms were revealed by clinical and immune characteristics, such as high expression of PD-L1, high tumor mutational burden (TMB-H), Epstein-Barr virus (EBV) positive and CD8+ tumor-infiltrating lymphocyte positive.

Evaluation of second-line apatinib plus irinotecan as a treatment for advanced gastric adenocarcinoma or gastroesophageal conjunction adenocarcinoma: a prospective, multicenter phase II trial.

Objective: Apatinib and irinotecan are used as systematic therapies for advanced gastric adenocarcinoma (GAC) and gastroesophageal junction adenocarcinoma (GEJA), while the evidence for their combination as second-line therapy in these patients is limited. This study aimed to evaluate the efficacy and safety of second-line apatinib plus irinotecan for the treatment of GAC and GEJA. Methods: In this prospective, multicenter phase II clinical study, 28 patients with advanced GAC or GEJA who received second-line apatinib plus irinotecan were recruited. Results: In total, 1 (3.6%) patient achieved complete response, 7 (25.0%) patients achieved partial response, 13 (46.4%) patients had stable disease, and 4 (14.3%) patients showed progressive disease, while clinical response was not evaluable or not assessed in 3 (10.7%) patients. The objective response rate and disease control rate were 28.6% and 75.0%, respectively. Meanwhile, the median (95% confidence interval (CI)) progression-free survival (PFS) was 4.5 (3.9-5.1) months, and the median (95% CI) overall survival (OS) was 11.3 (7.4-15.1) months. By multivariate Cox regression analysis, male sex, liver metastasis, and peritoneal metastasis were independently associated with worse PFS or OS, while treatment duration ≥5 months was independently associated with better OS. In terms of the safety profile, 89.3% of patients experienced treatment-emergent adverse events of any grade, among which 82.1% of patients had grade 1-2 adverse events and 64.3% of patients had grade 3-4 adverse events. Conclusion: Apatinib plus irinotecan as second-line therapy achieves a good treatment response and satisfactory survival with tolerable safety in patients with advanced GAC or GEJA.

Construction of the prognostic model for small-cell lung cancer based on inflammatory markers: A real-world study of 612 cases with eastern cooperative oncology group performance score 0-1.

OBJECTIVES: This research aimed to explore the relationship between pre-treatment inflammatory markers and other clinical characteristics and the survival of small-cell lung cancer (SCLC) patients who received first-line platinum-based treatment and to construct nomograms for predicting overall survival (OS) and progression-free survival (PFS). METHODS: A total of 612 patients diagnosed with SCLC between March 2008 and August 2021 were randomly divided into two cohorts: a training cohort (n = 459) and a validation cohort (n = 153). Inflammatory markers, clinicopathological factors, and follow-up information of patients were collected for each case. Cox regression was used to conduct univariate and multivariate analyses and the independent prognostic factors were adopted to develop the nomograms. Harrell's concordance index (C-index) and time-dependent receiver operating characteristic curve were used to verify model differentiation, calibration curve was used to verify consistency, and decision curve analysis was used to verify the clinical application value. RESULTS: Our results showed that baseline C-reactive protein/albumin ratio, neutrophil/lymphocyte ratio, NSE level, hyponatremia, the efficacy of first-line chemotherapy, and stage were independent prognostic factors for both OS and PFS in SCLC. In the training cohort, the C-index of PFS and OS was 0.698 and 0.666, respectively. In the validation cohort, the C-index of PFS and OS was 0.727 and 0.747, respectively. The nomograms showed good predictability and high clinical value. Also, our new clinical models were superior to the US Veterans Administration Lung Study Group (VALG) staging for predicting the prognosis of SCLC. CONCLUSIONS: The two prognostic nomograms of SCLC including inflammatory markers, VALG stage, and other clinicopathological factors had good predictive value and could individually assess the survival of patients.

TRIM27 is an adverse prognostic biomarker and associated with immune and molecular profiles in right-sided colon cancer.

Right-sided colon cancer (RCC), as an independent tumor entity, shows a poor prognosis. It is imperative to detect immune microenvironment-related genes for predicting RCC patient prognosis and study their function in RCC. Tripartite motif-containing 27 (TRIM27) was identified as a risk signature from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) datasets by using weighted gene co-expression network analysis, differentially expressed analysis, and univariate Cox analysis. It predicted a poorer overall survival and increased lymph node metastasis, which were then validated in our 48 clinical samples. Using immunohistochemistry, TRIM27 was found to be highly expressed in both cancer cells and surrounding immunocytes, and its expression in tumor or immune cells both predicted a poorer prognosis. Thereafter, the functional mechanism, immune and molecular characteristics of TRIM27 were investigated using gene set enrichment analysis (GSEA), ESTIMATE, CIBERSORT, and gene set variation analysis (GSVA) at the single-cell, somatic mutation, and RNA-seq level. Patients with highly expressed TRIM27 presented lower CD4+ T cell infiltration and activation of the mTORC1/glycolysis pathway. In addition, patients with highly expressed TRIM27 were characterized by hypermetabolism, higher tumor purity, more BRAF mutation, and more chromosomal instability. Collectively, TRIM27 is an important immune-related prognostic biomarker in patients with RCC. It may function via activating the mTORC1/glycolysis pathway and suppressing CD4+ T cells. These results indicated that TRIM27 could be a promising therapeutic target in RCC.

SOX8 promotes cetuximab resistance via HGF/MET bypass pathway activation in colorectal cancer.

AIM: Cetuximab is an essential drug for the treatment of wild-type K-RAS colorectal cancer (CRC). It improves the overall survival of patients. However, acquired resistance prevents its clinical efficacy. Tumor heterogeneity may be a nonnegligible reason for cetuximab resistance. We attempted to explore the corresponding molecular mechanism. METHODS: Cetuximab-resistant CRC cell RKO and cetuximab-sensitive CRC cell Caco-2 were applied in this study. Cells were centrifuged to determine the concentration in the culture supernatant (CS). MTT, EdU, and colony formation assays were utilized to evaluate cell survival and proliferation. Chromatin immunoprecipitation (ChIP) and promoter-luciferase reporter assays were employed to confirm the direct binding of transcription factors. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) assays were used to detect the expression of molecular markers in the pathway. RESULTS: Hepatocyte growth factor (HGF) was up-regulated in RKO cell culture supernatant and induced cetuximab resistance in Caco-2 cells. SRY-Box Transcription Factor 8 (SOX8) bound to the promoter sequence of HGF. HGF activated the HGF/MET bypass pathway and induced cetuximab resistance in Caco-2 cells. CONCLUSION: The SOX8/HGF/MET axis played a crucial role in the communication between cetuximab-resistant cells and cetuximab-sensitive cells, inducing treatment resistance.

Complete Pathologic Response of Multiple Liver Metastases and Clinical Complete Response of Rectal Cancer in a Patient with Ataxia-Telangiectasia Mutated Gene Mutations After XELOXIRI Plus Bevacizumab: A Case Report.

BACKGROUND: Doublet or triplet chemotherapy plus or minus targeted drugs can achieve a high objective response rate (ORR) and are currently considered to be the backbone of conventional therapy for liver metastatic colorectal cancer (mCRC). However, current biomarkers (such as UGT1A1 and DPYD) are limited to the prediction of toxicity and there are no effective biomarkers to predict chemotherapy response. Therefore, personalized cancer chemotherapy underpinned by genomic alterations in mCRC has received increasing attention. CASE PRESENTATION: We present a case of a 28-year-old female rectal cancer patient with multiple liver metastases (clinical risk score, CRS = 5 points). The patient underwent XELOXIRI plus bevacizumab regimen that consisted of irinotecan (150 mg/m2), oxaliplatin (100 mg/m2) on day 1, capecitabine (1700 mg/m2 per day from day 2 to 15), bevacizumab (7.5 mg/kg) on day 1 (on the second cycle), given every three weeks for eight cycles. After multi-disciplinary team (MDT) discussion, the patient underwent right hemihepatectomy, partial liver resection of segment IV and cholecystectomy. Surprisingly, the patient achieved a complete pathologic response (pCR) of the hepatic metastasis and clinical complete response (cCR) of the primary rectal lesion. A paired tumor molecular profile revealed somatic mutations in ataxia-telangiectasia mutated (ATM) genes that may explain why the patient achieved such a dramatic tumor response. Treatment was discontinued after eight cycles of a single oral dose of capecitabine and the patient started a follow-up program of physical and radiological examinations. To monitor the signs of recurrence, we also obtained blood samples to analyze circulating tumor DNA (ctDNA). To date, the patient has remained disease-free. CONCLUSION: The XELOXIRI-bevacizumab regimen is a feasible and effective regimen for patients with mCRC. Mutations in the ATM genes may characterize a subset of patients with a better prognosis who are more sensitive to chemotherapy plus biological agents.

A Prognostic Nomogram of Colon Cancer With Liver Metastasis: A Study of the US SEER Database and a Chinese Cohort.

BACKGROUND: Among colon cancer patients, liver metastasis is a commonly deadly phenomenon, but there are few prognostic models for these patients. METHODS: The clinicopathologic data of colon cancer with liver metastasis (CCLM) patients were downloaded from the Surveillance, Epidemiology and End Results (SEER) database. All patients were randomly divided into training and internal validation sets based on the ratio of 7:3. A prognostic nomogram was established with Cox analysis in the training set, which was validated by two independent validation sets. RESULTS: A total of 5,700 CCLM patients were included. Age, race, tumor size, tumor site, histological type, grade, AJCC N status, carcinoembryonic antigen (CEA), lung metastasis, bone metastasis, surgery, and chemotherapy were independently associated with the overall survival (OS) of CCLM in the training set, which were used to establish a nomogram. The AUCs of 1-, 2- and 3-year were higher than or equal to 0.700 in the training, internal validation, and external validation sets, indicating the favorable effects of our nomogram. Besides, whether in overall or subgroup analysis, the risk score calculated by this nomogram can divide CCLM patients into high-, middle- and low-risk groups, which suggested that the nomogram can significantly determine patients with different prognosis and is suitable for different patients. CONCLUSION: Higher age, the race of black, larger tumor size, higher grade, histological type of mucinous adenocarcinoma and signet ring cell carcinoma, higher N stage, RCC, lung metastasis, bone metastasis, without surgery, without chemotherapy, and elevated CEA were independently associated with poor prognosis of CCLM patients. A nomogram incorporating the above variables could accurately predict the prognosis of CCLM.

FEN1 is a prognostic biomarker for ER+ breast cancer and associated with tamoxifen resistance through the ERα/cyclin D1/Rb axis.

BACKGROUND: Tamoxifen is an important choice in endocrine therapy for patients with oestrogen receptor-positive (ER+) breast cancer, and disease progression-associated resistance to tamoxifen therapy is still challenging. Flap endonuclease-1 (FEN1) is used as a prognostic biomarker and is considered to participate in proliferation, migration, and drug resistance in multiple cancers, especially breast cancer, but the prognostic function of FEN1 in ER+ breast cancer, and whether FEN1 is related to tamoxifen resistance or not, remain to be explored. METHODS: On-line database Kaplan-Meier (KM) plotter, GEO datasets, and immunohistochemistry were used to analyse the prognostic value of FEN1 in ER+ breast cancer from mRNA and protein levels. Cell viability assay and colony formation assays showed the response of tamoxifen in MCF-7 and T47D cells. Microarray data with FEN1 siRNA versus control group in MCF-7 cells were analysed by Gene Set Enrichment Analysis (GSEA). The protein levels downstream of FEN1 were detected by western blot assay. RESULTS: ER+ breast cancer patients who received tamoxifen for adjuvant endocrine therapy with poor prognosis showed a high expression of FEN1. MCF-7 and T47D appeared resistant to tamoxifen after FEN1 over-expression and increased sensitivity to tamoxifen after FEN1 knockdown. Importantly, FEN1 over-expression could activate tamoxifen resistance through the ERα/cyclin D1/Rb axis. CONCLUSIONS: As a biomarker of tamoxifen effectiveness, FEN1 participates in tamoxifen resistance through ERα/cyclin D1/Rb axis. In the future, reversing tamoxifen resistance by knocking-down FEN1 or by way of action as a small molecular inhibitor of FEN1 warrants further investigation.

Biological and clinical significance of flap endonuclease­1 in triple­negative breast cancer: Support of metastasis and a poor prognosis.

Flap endonuclease­1 (FEN1), a structure­specific nuclease participating in DNA replication and repair processes, has been confirmed to promote the proliferation and drug resistance of tumor cells. However, the biological functions of FEN1 in cancer cell migration and invasion have not been defined. In the present study, using online database analysis and immunohistochemistry of the specimens, it was found that FEN1 expression was associated with a highly invasive triple­negative breast cancer (TNBC) subtype in both breast cancer samples from the Oncomine database and from patients recruited into the study. Furthermore, FEN1 was an important biomarker of lymph node metastasis and poor prognosis in patients with TNBC. FEN1 promoted migration of TNBC cell lines and FEN1 knockdown reduced the number of spontaneous lung metastasis in vivo. Ingenuity Pathway Analysis of FEN1­related transcripts in 198 patients with TNBC demonstrated that the polo­like kinase family may be the downstream target of FEN1. PLK4 was further identified as a critical target of FEN1 mediating TNBC cell migration, by regulating actin cytoskeleton rearrangement. The results of the present study validate FEN1 as a therapeutic target in patients with TNBC and revealed a new role for FEN1 in regulating TNBC invasion and metastasis.

Clinical outcomes of capecitabine-based versus S-1-based regimens as first-line chemotherapy in patients with unresectable or metastatic gastric cancer: a propensity score matched single-center comparison.

BACKGROUND: Fluoropyrimidine-based regimens are the cornerstone of first-line chemotherapy for metastatic gastric cancer (GC). Capecitabine or S-1 might be used as an alternative to infusional 5-fluorouracil, especially in pan-Asian. This study aimed to compare the clinical outcomes of capecitabine-based and S-1-based regimens as first-line chemotherapy in Chinese patients with unresectable or metastatic GC. METHODS: We conducted a retrospective study including unresectable or metastatic GC patients treated with the capecitabine-based or S-1-based regimen as first-line chemotherapy at the First Hospital of China Medical University. Propensity score matching (PSM) analysis was performed to reduce selection bias. Overall survival (OS) outcomes were compared using the Kaplan-Meier method and log-rank test. Prognostic significance was determined using multivariate Cox regression analysis. In addition, subgroup analyses were conducted to determine the effectiveness of capecitabine-based and S-1-based regimens in clinically relevant patient subsets. RESULTS: The clinical data of 469 patients included between October 2005 and September 2018. PSM analysis identified 187 patients receiving capecitabine-based or S-1-based regimen. No significant difference in OS (10.7 vs. 11.1 months, P=0.523) was detected between the two groups after PSM. In the subgroup analysis, the median OS (12.2 vs. 9.3 months, P=0.013) was longer for patients with peritoneum metastasis who received the capecitabine-based regimen compared to those who received the S-1-based regimen. CONCLUSIONS: No significant difference in clinical outcomes was observed between the capecitabine and S-1-based regimen as first-line chemotherapy for metastatic or unresectable GC patients in China. The capecitabine-based regimen should be considered in the treatment of the GC patients with peritoneum metastasis.

Nomogram-based prediction of survival in unresectable or metastatic gastric cancer patients with good performance status who received first-line chemotherapy.

BACKGROUND: Good performance status (PS) is widely acknowledged to have a high prognostic ability, although the prognostic parameters of cancer patients with good PS are still uncertain. This study was conducted to establish and validate a point-based nomogram to assist with predicting prognosis in unresectable or metastatic gastric cancer (GC) patients who had good PS and underwent first-line chemotherapy. METHODS: At random, a total of 309 patients with GC were split into 2 cohorts: a training cohort (n=259) and an internal validation cohort (n=50). An independent external validation cohort comprising 147 patients was also recruited. Both univariate and multivariate Cox regression analyses were used to evaluate patients based on the overall survival (OS) to develop the nomogram, which was subsequently validated using the concordance index (c-index), calibration curve, and decision curve analysis (DCA). RESULTS: The nomogram contained 3 independent prognostic variables in the training cohort: the number of distant metastatic sites (P<0.001), carbohydrate antigen 199 (CA199) level (P=0.002), and fibrinogen (P=0.020). The nomogram predicted an OS with a c-index of 0.623 (95% CI, 0.58-0.67) in the training cohort. The internal validation showed that the nomogram had a c-index of 0.614 (95% CI, 0.51-0.72). For external validation, the c-index was 0.638 (95% CI, 0.58-0.70). CONCLUSIONS: A reliable point-based nomogram for predicting the prognosis of patients who had unresectable or metastatic GC and good PS who underwent first-line chemotherapy was developed and validated. KEYWORDS: Nomogram-based prediction; overall survival; unresectable gastric cancer; metastatic gastric cancer; good performance status; first-line chemotherapy.

Prediction of KRAS, NRAS and BRAF status in colorectal cancer patients with liver metastasis using a deep artificial neural network based on radiomics and semantic features.

There is a critical need for development of improved methods capable of accurately predicting the RAS (KRAS and NRAS) and BRAF gene mutation status in patients with advanced colorectal cancer (CRC). The purpose of this study was to investigate whether radiomics and/or semantic features could improve the detection accuracy of RAS/BRAF gene mutation status in patients with colorectal liver metastasis (CRLM). In this retrospective study, 159 patients who had been diagnosed with CRLM in two hospitals were enrolled. All patients received lung and abdominal contrast-enhanced CT (CECT) scans prior to radiation therapy and chemotherapy. Semantic features were independently assessed by two radiologists. Radiomics features were extracted from the portal venous phase (PVP) of the CT scan for each patient. Seven machine learning algorithms were used to establish three scores based on the semantic, radiomics and the combination of both features. Two semantic and 851 radiomics features were used to predict the mutation status of RAS and BRAF using an artificial neural network method (ANN). This approach performed best out of the seven tested algorithms. We constructed three scores which were based on radiomics, semantic features and the combined scores. The combined score could distinguish between wild-type and mutant patients with an AUC of 0.95 in the primary cohort and 0.79 in the validation cohort. This study proved that the application of radiomics together with semantic features can improve non-invasive assessment of the gene mutation status of RAS (KRAS and NRAS) and BRAF in CRLM.

Exosomal PD-L1 Retains Immunosuppressive Activity and is Associated with Gastric Cancer Prognosis.

BACKGROUND: A recent study showed that circulating exosomal PD-L1 is an effective predictor for anti-PD-1 therapy in melanomas. Exosomal PD-L1 induced immunosuppression microenvironments in cancer patients. However, its prognostic value and immunosuppressive effect in gastric cancer (GC) were poorly understood. METHODS: We retrospectively evaluated the prognostic value of exosomal PD-L1 and soluble PD-L1 in preoperative plasma of 69 GC patients. The correlation between exosomal PD-L1 and the T cell counts or cytokine in the plasma was evaluated in 31 metastatic GC patients before chemotherapy. RESULTS: Overall survival (OS) was significantly lower in the high exosomal PD-L1 group compared with the low exosomal PD-L1 group (P = 0.004). Exosomal PD-L1 was an independent prognostic factor in GC (n = 69, 95% confidence interval = 1.142-7.669, P = 0.026). However, soluble PD-L1 showed no correlation with OS (P = 0.139). Additionally, exosomal PD-L1 in the plasma samples of 31 metastatic GC patients was negatively associated with CD4+ T cell count (P = 0.001, R = - 0.549), CD8+ T-cell count (P = 0.054, R = - 0.349), and granzyme B (P = 0.002, R = - 0.537), indicating that exosomal PD-L1 was associated with immunosuppressive status of GC patients. GC cells also secreted exosomal PD-L1 and were positively associated with the amount of PD-L1 in corresponding GC cell lines. Besides, exosomal PD-L1 significantly decreased T-cell surface CD69 and PD-1 expressions compared with soluble PD-L1 due to its stable and MHC-I expression. CONCLUSIONS: Overall, exosomal PD-L1 predicts the worse survival and reflects the immune status in GC patients, resulting from a stronger T-cell dysfunction due to its stable and MHC-I expression.

The dynamic monitoring of CEA in response to chemotherapy and prognosis of mCRC patients.

BACKGROUND: The role of carcinoembryonic antigen (CEA) change patterns in tumor response and long-term outcome is unclear. This study aimed to investigate the correlation between changes in CEA levels and tumor response as a potential prognostic model. METHODS: CEA levels were determined from baseline to progression. A χ2 test was used to assess the correlation between CEA changes and tumor response. Univariate and multivariate COX models were used to explore the correlation of CEA changes to progression-free survival (PFS) and overall survival (OS). RESULTS: All 114 patients were divided into five groups according to CEA change pattern (A: patients had an initial fast CEA decrease that then turned into a slow increase; B: patients had an initial slow CEA decrease that then turned to a slow increase; C: patients had a continually slow CEA increase; D: patients had a continually fast CEA increase; E: patients had an initial fast CEA decrease that then turned into a fast increase). Patients in Group A had the longest OS and PFS while Group E patients had the shortest OS. Baseline to week 12 and week 12 to week 18 change rates were consistent with tumor response and progression, respectively. An increase in CEA level by ≥2.7% from week 12 to 18 was an independent negative prognostic factor of OS. CONCLUSIONS: CEA changes mirror the tumor response to first-line chemotherapy and are associated with prognosis. CEA monitoring may be a substitute for computed tomography during the CEA stable period of treatment.

Caveolin­1 enhances RANKL­induced gastric cancer cell migration.

The classical pathway involving receptor activator of nuclear factor­κB (RANK) and its ligand (RANKL) induces the activation of osteoclasts and the migration of a variety of tumor cells, including breast and lung cancer. In our previous study, the expression of RANK was identified on the surface of gastric cancer cells, however, whether the RANKL/RANK pathway is involved in the regulation of gastric cancer cell migration remains to be fully elucidated. Lipid rafts represent a major platform for the regulation of cancer signaling; however, their involvement in RANKL­induced migration remains to be elucidated. To investigate the potential roles and mechanism of RANKL/RANK in gastric cancer migration and metastasis, the present study examined the expression of RANK by western blot analysis and the expression of caveolin­1 (Cav­1) in gastric cancer tissues by immunohistochemistry, in addition to cell migration which is measured by Transwell migration assay. The aggregation of lipid reft was observed by fluorescence microscopy and western blotting was used to measure signaling changes in associated pathways. The results showed that RANKL induced gastric cancer cell migration, accompanied by the activation of Cav­1 and aggregation of lipid rafts. Nystatin, a lipid raft inhibitor, inhibited the activation of Cav­1 and markedly reversed RANKL­induced gastric cancer cell migration. The RANKL­induced activation of Cav­1 has been shown to occur with the activation of proto­oncogene tyrosine­protein kinase Src (c­Src). The c­Src inhibitor, PP2, inhibited the activation of Cav­1 and lipid raft aggregation, and reversed RANKL­induced gastric cancer cell migration. Furthermore, it was demonstrated that Cav­1 was involved in RANKL­induced cell migration in lung, renal and breast cancer cells. These results suggested that RANKL induced gastric cancer cell migration, likely through mechanisms involving the c­Src/Cav­1 pathway and lipid raft aggregation.

MicroRNA-29b-2-5p inhibits cell proliferation by directly targeting Cbl-b in pancreatic ductal adenocarcinoma.

BACKGROUND: MicroRNAs can be used in the prognosis of malignancies; however, their regulatory mechanisms are unknown, especially in pancreatic ductal adenocarcinoma (PDAC). METHODS: In 120 PDAC specimens, miRNA levels were assessed by quantitative real time polymerase chain reaction (qRT-PCR). Then, the role of miR-29b-2-5p in cell proliferation was evaluated both in vitro (Trypan blue staining and cell cycle analysis in the two PDAC cell lines SW1990 and Capan-2) and in vivo using a xenograft mouse model. Next, bioinformatics methods, a luciferase reporter assay, Western blot, and immunohistochemistry (IHC) were applied to assess the biological effects of Cbl-b inhibition by miR-29b-2-5p. Moreover, the relationship between Cbl-b and p53 was evaluated by immunoprecipitation (IP), Western blot, and immunofluorescence. RESULTS: From the 120 PDAC patients who underwent surgical resection, ten patients with longest survival and ten with shortest survival were selected. We found that high miR-29b-2-5p expression was associated with good prognosis (p = 0.02). The validation cohort confirmed miR-29b-2-5p as an independent prognostic factor in PDAC (n = 100, 95% CI = 0.305-0.756, p = 0.002). Furthermore, miR-29b-2-5p inhibited cell proliferation, induced cell cycle arrest, and promoted apoptosis both in vivo and in vitro. Interestingly, miR-29b-2-5p directly bound the Cbl-b gene, down-regulating its expression and reducing Cbl-b-mediated degradation of p53. Meanwhile, miR-29b-2-5p expression was negatively correlated with Cbl-b in PDAC tissues (r = - 0.33, p = 0.001). CONCLUSIONS: Taken together, these findings indicated that miR-29b-2-5p improves prognosis in PDAC by targeting Cbl-b to promote p53 expression, and would constitute an important prognostic factor in PDAC.

The Chemokine Receptor CXCR4 and c-MET Cooperatively Promote Epithelial-Mesenchymal Transition in Gastric Cancer Cells.

The C-X-C motif chemokine receptor 4 (CXCR4) pathway can promote tumor metastasis but is dependent on cross talk with other signaling pathways. The MET proto-oncogene (c-MET) participates in metastasis and is highly expressed in gastric cancer. However, the relationship between CXCR4 and c-MET signaling and their mechanisms of action in gastric cancer metastasis remain unclear. In this study, in vitro experiments demonstrated that C-X-C motif chemokine ligand 12 (CXCL12)/CXCR4 induces epithelial-mesenchymal transition (EMT) and promotes migration in gastric cancer cells, which is accompanied by c-MET activation. These phenomena were reversed by c-MET inhibition. Further investigation revealed that c-MET activation correlated with its interaction with caveolin 1 in lipid rafts, induced by CXCL12. In clinical samples, we observed a significant positive association between CXCR4 expression and c-MET phosphorylation (r = 0.259, P = .005). Moreover, samples expressing both receptors were found to indicate significantly poorer patient prognosis (P < .001). These results suggest that CXCL12 induces EMT at least partially through cross talk between CXCR4 and c-MET signaling. In addition, changes in these pathways could have clinical importance for the treatment of gastric cancer.

Pretreatment platelet-to-lymphocyte ratio is associated with the response to first-line chemotherapy and survival in patients with metastatic gastric cancer.

BACKGROUND: Several studies have shown that platelet-to-lymphocyte ratio (PLR) is a prognostic factor for various cancers. However, there is no study about the role of PLR in predicting response to first-line chemotherapy of metastatic gastric cancer. Therefore, this study aimed to establish whether PLR is associated with the response to first-line chemotherapy and survival in patients with metastatic gastric cancer. METHODS: We enrolled 273 patients diagnosed with metastatic gastric cancer. The best cut-off value of PLR to predict chemotherapeutic response was chosen by receiver operating characteristic (ROC) curve analysis. Prognostic significance was determined using the log-rank test and multivariate Cox regression analysis. RESULTS: Based on the cut-off value of PLR, patients were divided into a low PLR group and high PLR group. In logistic regression analysis, the low PLR group had a significantly higher disease control rate than the high PLR group had (91.3 vs 76.1%, P=.002), and PLR was an independent risk factor for response to first-line chemotherapy (odds ratio [OR]: 3.256; 95% confidence interval [CI]: 1.521-6.969; P=.002). The low PLR group had significantly longer overall survival (OS) than the high PLR group had (13.4 vs 9.2 months; P=.020). Multivariate survival analysis showed that PLR was significantly associated with OS [hazard ratio (HR): 1.002; 95% CI: 1.000-1.003; P=.020]. CONCLUSIONS: Pre-treatment PLR is associated with the response rate to first-line chemotherapy and survival outcomes in patients with metastatic gastric cancer.

Combination of platelet count and neutrophil-lymphocyte ratio as a prognostic marker to predict chemotherapeutic response and survival in metastatic advanced gastric cancer.

AIM: The study evaluated the prognostic impact of combination of platelet count and neutrophil-lymphocyte ratio (COP-NLR) for first-line chemotherapeutic response and survival outcomes in metastatic advanced gastric cancer patients. METHODS: Two hundred and seventy-three patients were categorized into three COP-NLR groups (COP-NLR 0, 1, 2) according to their platelet count and neutrophil-lymphocyte ratio. RESULTS: The COP-NLR 0 had a significantly higher disease control rate (93%) than the other two groups (p = 0.011). A logistic regression model showed that COP-NLR was an independent risk factor for response to chemotherapy (odds ratio: 2.247; 95% CI: 1.303-3.874; p = 0.044). The median overall survival for COP-NLR 0, 1 and 2 was 14.8, 10.3 and 9.1 months, respectively (p = 0.001). CONCLUSION: COP-NLR is a useful predictor of survival outcomes and chemotherapeutic response in patients with metastatic advanced gastric cancer.