Relationship Between MUC4 Variants and Metastatic Recurrence in Colorectal Cancer.

Background: Recurrent metastasis after radical resection in patients of colorectal cancer (CRC) is a great challenge for the world, in which genomic alterations play a major role in tumorigenesis. MUC4 plays a significant role in recurrence and metastasis in tumor. This study is aimed at exploring the association between MUC4 variants and metastatic recurrence of CRC. Methods: Forty-seven patients relapsing with metastasis and 37 patients remaining disease-free postoperatively were enrolled. Next-generation sequencing (NGS) detected mutations. Mutation and mRNA expression data were downloaded from TCGA and cBioPortal databases. We analyzed the relationship between MUC4 variants and clinical parameters, as well as possible molecular mechanisms. Results: MUC4 variants rs56359992 and rs781124621 were associated with survival in patients with CRC. Rs56359992 was more common in patients with metastatic recurrence. MAPK pathway, PI3K-Akt pathway, JAK-STAT pathway, cell cycle, WNT pathway and mTOR pathway were found to correlate with MUC4 mutation by GO/KEGG analysis, as well as resting and activated mast cell related to MUC4 mutation by CIBERSORT analysis. Conclusion: Genetic variants of MUC4 with CRC may constitute a molecular signature of metastatic recurrence. MUC4 may become a new target for the treatment of CRC recurrence.

Efficacy of Camrelizumab in Advanced Non-Small-Cell Lung Cancer and Prognostic Analysis of Different PET/CT Features.

Objective: To evaluate the efficacy of the PD-1 inhibitor camrelizumab plus chemotherapy in the first-line treatment of advanced non-small-cell lung cancer (NSCLC) and the prognostic differences of patients with different PET/CT features. Methods: Between December 2018 and October 2020, 100 patients with NSCLC assessed for eligibility treated in our institution were recruited and randomly assigned (1 : 1) to receive either the TC regimen chemotherapy (control group) or the TC regimen chemotherapy plus camrelizumab (study group). The primary endpoints were clinical efficacy, progression-free survival (PFS), and overall survival (OS). A decrease of max standard uptake value (SUVmax) of >30% in primary lung cancer was considered as metabolic remission. The prognostic differences of the eligible patients with different PET/CT features were assessed. Survival data were analyzed using the Kaplan-Meier method to obtain the survival rate and calculate the median survival time. Results: The metabolic remission rate and objective remission rate were significantly higher with chemotherapy plus camrelizumab versus chemotherapy alone. The study group had significantly higher CD3+ and CD4+ T-cell ratios and CD4+/CD8+ ratio and significantly lower CD8+ T-cell ratio than the control group after treatment. PFS (10 months versus 4 months) and OS (HR = 37.094, P ≤ 0.001) were better with camrelizumab plus chemotherapy versus stand-alone chemotherapy. The incidence of adverse events (AE) was similar between the two groups. The patients in the study group were stratified into metabolic remission and metabolic nonremission based on PET/CT results. Intersubgroup analysis showed significantly better PFS and OS in the metabolic remission group than in the nonmetabolic remission group. Conclusion: The camrelizumab plus chemotherapy as a first-line treatment option for NSCLC significantly increases the survival benefit. Metabolic status shown by PET/CT correlates with long-term prognosis and demonstrates a great potential for early assessment of efficacy to support the choice of treatment regimens.

High expression of EZR (ezrin) gene is correlated with the poor overall survival of breast cancer patients.

BACKGROUND: To evaluate the EZR (ezrin) gene expression in breast cancer and correlation with the prognosis through bioinformatics analysis and immunohistochemistry assay. METHODS: EZR gene expression in breast cancer and corresponding normal breast tissue was compared in the TCGA database. Protein-protein interaction (PPI) network relevant EZR was established through the STRING database. The correlation between EZR expression and prognosis of breast cancer was analyzed by the log-rank analysis from the TCGA. Ezrin protein (coded by EZR) expression was also examined by immunohistochemistry assay in 120 breast cancer patients. RESULTS: EZR expression level in tumor tissue was significantly upregulated compared to that of normal breast tissue of breast cancer patients (P < 0.05). In the PPI analysis, there were 51 nodes and 455 edges in the network. The top 10 hub genes of the network were identified. High expression of EZR mRNA was correlated with poor overall survival (OS) of the breast cancer patients (HR = 1.40, P = 0.038). However, the disease-free survival (DFS) of breast cancer patients did not correlate with the EZR mRNA level (HR = 0.86, P = 0.44). The ezrin protein expression was positive with uniform brown-yellow granules in the cell membrane, cavity surface and cytoplasm of the breast cancer cells. Of the included 120 cancer samples, 98 cases were positive for ezrin expression and 22 were negative. No correlation was found between ezrin expression site and patients' clinicopathological features. CONCLUSION: EZR is upregulated in breast cancer and can be used as potential biomarker for overall survival.

miR-30a inhibits breast cancer progression through the Wnt/ß-catenin pathway.

BACKGROUND: miR-30a is a microRNA associated with the progression of malignant tumors such as gastric cancer, colon cancer, prostate cancer, and lung cancer, and can regulate the proliferation and migration of breast cancer (BC) cells in vitro. However, its expression, function, clinical significance and relationship with the Wnt/ß-catenin pathway in human BC were still unclear. METHODS: Immunohistochemistry, Western blotting and real-time quantitative PCR (RT-qPCR) were used to measure the expressions of miR-30a and ß-catenin in 114 pairs of human BC tumor tissues and adjacent normal tissues which were collected from March 2014 to October 2015. The effect of miR-30a on the expression of ß-catenin was studied in the MCF-7 cells in vitro. RESULTS: The expression levels of miR-30a in human BC tumor tissues were significantly lower than they were in the adjacent normal tissues (P < 0.001), and significantly higher in ß-catenin protein (P < 0.001), but there was no significant different in ß-catenin mRNA (P = 0.3816). The immunohistochemistry results showed that ß-catenin protein was only expressed on the cell membrane in paracancerous normal tissues, but ß-catenin protein was expressed on the cell membrane and cytoplasm in BC tumor cells. In addition, there was a significantly negative correlation (r = -0.816, P < 0.001) between the expression miR-30a and ß-catenin protein in BC tissues. The age of onset, PR expression, ER expression, and HER-2 expression of the BC patients were not related to miR-30a or ß-catenin protein expression (P > 0.05). Tumor diameter, histological grade, lymph node metastasis, TNM stage, and the prognosis of BC patients (P < 0.05) were significantly related to miR-30a or ß-catenin protein expression. In MCF-7 cells, miR-30a regulated the accumulation of ß-catenin protein by inhibiting the expression of BCL9 in BC cells. CONCLUSION: miR-30a was lowly expressed in breast cancer tissues and highly in ß-catenin protein, and miR-30a might block the Wnt/ß-catenin pathway by inhibiting the accumulation of ß-catenin, and then inhibiting breast cancer progression.

Novel long noncoding RNA GACAT3 promotes colorectal cancer cell proliferation, invasion, and migration through miR-149.

AIM: To explore the expression and clinical significance of long noncoding RNA (lncRNA) gastric cancer-associated transcript 3 (GACAT3) in human colorectal cancer (CRC). METHODS: Expression of GACAT3 in CRC tissues and cell lines was measured using quantitative real-time PCR. CCK-8 and colony formation assays were used to assess the effect of GACAT3 on CRC cell line proliferation. Transwell invasion and migration assays were performed to detect the effect of GACAT3 on CRC cell line invasion and migration. Bioinformatics prediction, luciferase reporter assay, and pull-down assay were used to determine if miR-149 was a target of GACAT3. In addition, we also conducted colony formation assays and invasion assays to verify that GACAT3 promotes tumor progression through miR-149. Finally, in vivo tumorigenesis studies were used to demonstrate subcutaneous tumor growth. RESULTS: In the present study, we found that GACAT3 was highly expressed in CRC tissues and cell lines. Si-GACAT3 significantly decreased cell proliferation, motility, and invasiveness both in vitro and in vivo. We confirmed that downregulated GACAT3 significantly increased the expression of miR-149, and miR-149 binds to GACAT3 in a sequence-specific manner using luciferase reporter assays and pull-down assay. Further functional experiments indicated that GACAT3 could directly upregulate SP1 and STAT3 expressions by functioning as a competing endogenous RNA for miR-149, and consequentially, promoting CRC cell proliferation and invasion in vitro. CONCLUSION: This study demonstrated that GACAT3 promotes tumor progression through competitive binding to miR-149 and suggests a promising new strategy for anti-CRC therapy.

A sensitive chemiluminescent immunoassay to detect Chromotrope FB (Chr FB) in foods.

Chromotrope FB (Chr FB) is a synthetic azo dye permitted for use in foods and medicines. An acceptable daily intake (ADI) of Chr FB was 0-0.5mg/kg in China. In this study, we synthesized a Chr FB hapten with an amino group to prepare its artificial immunogen. Polyclonal antibodies obtained from New Zealand rabbits were applied to develop an indirect competitive chemiluminescent immunoassay (icCLIA) to detect Chr FB in foods. A horseradish peroxidase (HRP)-luminol-H2O2 system was used to yield CL signal with p-iodophenol as an enhancement reagent. The method showed good specificity towards Chr FB and could detect as low as 0.02ngmL-1 Chr FB in buffer, 0.07ngg-1 in yoghurt candy, 0.07ngg-1 in vitamin drink and 0.13ngg-1 in bread. Compared with HPLC method, the proposed method is more sensitive by two orders of magnitude. The accuracy and precision of this method are acceptable and comparable with HPLC method. Therefore, the proposed method could be used for rapid screening of Chr FB in the mentioned foodstuffs.

Relationship between the expression of TRß1 and the molecular typing and clinicopathological features of breast cancer.

OBJECTIVE: To study the relationship between the expression of TRß1 and the molecular typing and clinicopathological features of breast cancer. METHODS: The expression of TRß1, ER, PR and HER-2 proteins in 208 cases of invasive breast cancer, 52 intraductal carcinoma and 22 normal breast tissue was detected by immunohistochemistry in order to analyze the relationship between the expression of TRß1 protein and clinicopathological parameters of breast cancer. Western blot was performed to detect the effect of TRß1 silencing on the expression of Notch signaling pathway proteins and Epithelial-mesenchymal transition (EMT)-related proteins in MCF-7 cells. RESULTS: In the 208 cases of invasive breast cancer tissues, over expression of TRß1 protein was found in 88 cases while low expression in 120 cases, and the immunohistochemical score was (3.9±3.1). TRß1 protein was found over expressed in all the 52 cases of intraductal carcinoma and 22 cases of normal breast tissue, with the immunohistochemical score of (9.7±2.1) and 12.0, respectively, and there was no significant difference between the two groups (P>0.05) while both of them were significantly lower than the invasive breast cancer group (P<0.05). The expression of TRß1 protein in the invasive breast cancer tissues was significantly correlated with lymph node metastasis (P=0.041), molecular typing (P=0.037) and histological grade (P<0.001) while it was negatively correlated with HER-2 expression (r=0.926; P<0.001) and irrelevant with age (P=1.024), ER expression (P=0.834), PR expression (P=0.351) or TNM staging (P=1.032). Compared to normal MCF-7 cells, the expression of Notch1, Dell 1, Jagged-1 and vimentin proteins increased by 1.44 times, 1.53 times, 1.50 times and 1.45 times respectively in the TRß1 expression silenced MCF-7 cell. CONCLUSIONS: The expression of TRß1 protein in breast cancer tissues decreased with the increase of HER-2 expression and histological grade. The depletion of TRß1 protein may activate the Notch signaling pathway and enhance the EMT ability of breast cancer cells thus promoting the cancer cell migration.

PI3K/Akt signaling is involved in the pathogenesis of bleomycin­induced pulmonary fibrosis via regulation of epithelial­mesenchymal transition.

Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease characterized by chronic inflammation, fibroblast proliferation and extracellular matrix deposition. However, the molecular and cellular mechanisms underlying the pathogenesis of pulmonary fibrosis remain to be fully elucidated. The contribution of the phosphoinositide 3­kinase (PI3K)/protein kinase B (Akt) pathway in fibrotic processes remains to be investigated. The aim of the present study was to investigate the role of the PI3K/Akt pathway in pulmonary fibrosis. A rat model of pulmonary fibrosis was induced by intratracheal administration of bleomycin (BLM), and a specific PI3K/Akt inhibitor, LY294002, was used to assess the role of the PI3K/Akt pathway in fibrogenesis. The inflammatory and fibrotic alterations in the lung tissues were evaluated using histological staining and the hydroxyproline assay. In addition, the concentration of cytokines in bronchoalveolar lavage fluid and the expression of Akt, phosphorylated (p­)Akt, epithelial cadherin, α smooth muscle actin and vimentin in lung tissues. The data demonstrated that an increase in the expression levels of p­Akt was involved in the progression of pulmonary fibrosis and contributed to fibrogenesis. Administration of the Akt inhibitor significantly attenuated inflammation and fibrosis, which was accompanied by a reversal of lung fibrosis­associated epithelial­mesenchymal transition. Taken together, these observations suggest that the PI3K/Akt pathway serves a central role in the pathophysiology of lung fibrosis, and is a promising therapeutic target.

Preliminary study on CT perfusion imaging in guiding biopsy of pulmonary lumps.

BACKGROUND: CT perfusion imaging (CTP) has been proved to be a powerful functional imaging technique. This study aimed to evaluate the value of CTP in guiding biopsy of pulmonary lumps. METHODS: A total of 147 patients with pulmonary lumps who had CT guided biopsies were enrolled in this study from February 2005 to June 2007. The patients were assigned to 3 groups: 33 cases guided by CTP as group I, 45 cases guided by contrast-enhanced scan of CT as group II, and 69 cases guided by plain scan of CT as group III. Each group was subdivided into central and peripheral types according to the location of the lumps. The achievement ratio of biopsy, the accuracy in grouping, and grading of lung cancer, and the incidence of complication were compared. RESULTS: The total achievement ratios of biopsy from group I to III were 100% (33/33), 91% (41/45), and 80% (55/69) respectively, and the difference was statistically significant between group I and III (P < 0.05). For the central type, they were 100% (18/18), 88% (15/17), and 79% (11/14) respectively, and the difference was also statistically significant between group I and III (P < 0.05). For the peripheral type, they were 100% (15/15), 93% (26/28), and 80% (44/55) respectivelies, and the difference was not statistically significant among the three groups. The total accuracies in grouping and grading of lung cancer from group I to III were 100% (27/27), 91% (31/34), and 72% (33/46) respectively, and the difference was statistically significant between group I and III and between group II and III (P < 0.05). For the central type, they were 100% (16/16), 94% (16/17), and 70% (8/12) respectively, and the difference was statistically significant between group I and III (P < 0.05). For the peripheral type, they were 100% (11/11), 88% (15/17), and 72% (26/36) respectively, and the difference was statistically significant between group I and III (P < 0.05). The total incidence of complication from group I to III were 15% (5/33), 27% (12/45), and 43% (30/69) respectively, and the difference was statistically significant between group I and III (P < 0.01). For the central type, they were 11% (2/18), 24% (4/17), and 57% (8/14) respectively, and the difference was statistically significant between group I and III (P < 0.01). For the peripheral type, they were 20% (3/15), 29% (8/28), and 40% (22/55) respectively, and no statistically significant difference was found among the three groups. CONCLUSIONS: CTP guided biopsy of pulmonary lumps using multi-detector row CT has the potential to improve the accuracy of histopathological diagnosis with a lower risk and higher achievement ratio. More research and technical improvements are needed before it is widely used.