C-C Motif Chemokine Ligand 5 (CCL5) as a Predictive Marker for Non-sentinel Lymph Node Presence in Initial-Stage Breast Cancer with 1-2 Affected Sentinel Lymph Nodes.

The objective of our study was to use quantum dots for the purpose of seeing and detecting C-C motif chemokine ligand 5 (CCL5) inside the tissue of sentinel lymph nodes (SLN) and primary tumors. This endeavor aimed to enhance the accuracy of predicting the condition of non-sentinel lymph nodes and provide valuable insights for making informed treatment choices. We analyzed breast cancer patients who underwent sentinel lymph node biopsy followed by axillary lymph node removal due to one or two positive sentinel lymph nodes at the Second People's Hospital of Wuhu, China, between August 2018 and July 2022. Quantum dot technology was employed to visualize and determine CCL5 in the tissue samples from 84 patients. Out of a group diagnosed with breast cancer, 208 underwent sentinel lymph node biopsy. From this pool, 84 tested positive and subsequently underwent axillary lymph node removal. The presence of distinct orange-red fluorescence, linked to quantum dots, was evident in the cellular components of both primary tumors and positive sentinel lymph node tissues. We found a significant relationship between higher levels of SLNCCL5 and advanced tumor growth (P < 0.05). To understand the predictive value of SLN CCL5 related to non-sentinel lymph node status, we utilized the receiver operating characteristic (ROC) method. The area under the curve (AUC) calculated was 0.745 with a cutoff point of 23.285. Multivariate logistic regression was used to understand the effect of tumor dimensions and SLNCCL5 levels on non-sentinel lymph node status in specific patients. Both the size of the tumor and SLNCCL5 levels were found to have a significant impact (P < 0.05). Data suggested that the presence of positive SLNCCL5 might serve as an assessment parameter for anticipating the condition of non-SLN in cases of breast cancer involving T1 or T2 tumors with one or two positive sentinel lymph nodes.

Drug discovery by targeting the protein-protein interactions involved in autophagy.

Autophagy is a cellular process in which proteins and organelles are engulfed in autophagosomal vesicles and transported to the lysosome/vacuole for degradation. Protein-protein interactions (PPIs) play a crucial role at many stages of autophagy, which present formidable but attainable targets for autophagy regulation. Moreover, selective regulation of PPIs tends to have a lower risk in causing undesired off-target effects in the context of a complicated biological network. Thus, small-molecule regulators, including peptides and peptidomimetics, targeting the critical PPIs involved in autophagy provide a new opportunity for innovative drug discovery. This article provides general background knowledge of the critical PPIs involved in autophagy and reviews a range of successful attempts on discovering regulators targeting those PPIs. Successful strategies and existing limitations in this field are also discussed.

Combining ADC values in DWI with rCBF values in arterial spin labeling (ASL) for the diagnosis of mild cognitive impairment (MCI).

We aimed to investigate the role of combined apparent diffusion coefficient (ADC) values and relative cerebral blood flow (rCBF) values in the diagnosis of mild cognitive impairment (MCI) patients. The present prospective research enrolled 156 MCI patients and 58 healthy elderly people who came to our hospital from January 2021 to February 2023. T1W, T2W, diffusion-weighted imaging, and arterial spin labeling sequences were performed on all subjects, and ADC values and rCBF values were measured at the workstation. Clinical and demographic data of all patients were collected while mini-mental state examination (MMSE) and Montreal cognitive assessment (MoCA) scores were used to assess patients' cognitive abilities. The MCI group had significantly lower rCBF values in the left frontal lobe, left occipital lobe, right frontal lobe, and right occipital lobe than the HC group. The ADC values in the left frontal lobe as well as the right frontal lobe were remarkably elevated in the MCI group than in the HC group. MoCA and MMSE scores were positively correlated with rCBF values in the left frontal, right frontal, left occipital, and right occipital lobes and negatively correlated with ADC values in the left and right frontal lobes. Combined ADC values and rCBF values from the left frontal lobe for the diagnosis of MCI had a higher sensitivity and specificity with the AUC was 0.877, sensitivity 81.0%, specificity 82.7%. Additionally, pressure fasting plasma glucose, ADC of the left frontal lobe, right frontal lobe, rCBF of left frontal lobe and rCBF of left frontal lobe were the risk factors of patients with MCI. In summary, our results indicated that the ADC values and rCBF values were changed in MCI group compared to HC group and correlated with MMSE and MoCA scores.

Carcinosarcoma of common bile duct: A case report.

BACKGROUND: Carcinosarcomas of the common bile duct (CBD) are an extremely rare finding in the clinical setting. Based on a review of 12 literatures, 3 cases had the imaging features of ossification. Carcinosarcomas are prone to distant metastasis, as they possess clinical features of both carcinoma and sarcoma, and generally have with a poor prognosis. Due to the small number of cases reported, clinical experience in the diagnosis and treatment of the disease is lacking. CASE SUMMARY: The patient was a 75-year-old woman who had experienced recurrent chills with nausea and vomiting for 3 mo. Computed tomography, magnetic resonance imaging, endoscopic ultrasonography and endoscopic retrograde cholangiopancreatography led to the diagnosis of malignant tumor of the CBD. The patient ultimately underwent cholecystectomy, CBD resection, and choledochojejunostomy. Postoperative pathological examination revealed carcinosarcoma of the CBD, and the latest follow-up showed that the patient is recovering well. Based on previous case reports, some carcinosarcoma has ossification characteristics in imaging. If it is misdiagnosed as biliary calculi, the use of laser lithotripsy in surgery may lead to tumor diffusion. Choledochoscopy and narrow band staining of mucosa are very important for diagnosis. CONCLUSION: We herein present a rare case of carcinosarcomas of the CBD, we found the tumours may have imaging features of polypoid growth and ossification only when the sarcomal components are bone differentiation, while show soft tissue shadow when non bone differentiation. Confirmation of diagnosis depends greatly upon postoperative pathological examination and the adjuvant treatment has not been established, which leads to the poor prognosis.

Discovery of Small-Molecule Autophagy Inhibitors by Disrupting the Protein-Protein Interactions Involving Autophagy-Related 5.

One possible strategy for modulating autophagy is to disrupt the critical protein-protein interactions (PPIs) formed during this process. Our attention is on the autophagy-related 12 (ATG12)-autophagy-related 5 (ATG5)-autophagy-related 16-like 1 (ATG16L1) heterotrimer complex, which is responsible for ATG8 translocation from ATG3 to phosphatidylethanolamine. In this work, we discovered a compound with an (E)-3-(2-furanylmethylene)-2-pyrrolidinone core moiety (T1742) that blocked the ATG5-ATG16L1 and ATG5-TECAIR interactions in the in vitro binding assay (IC50 = 1-2 µM) and also exhibited autophagy inhibition in cellular assays. The possible binding mode of T1742 to ATG5 was predicted through molecular modeling, and a batch of derivatives sharing essentially the same core moiety were synthesized and tested. The outcomes of the in vitro binding assay and the flow cytometry assay of those newly synthesized compounds were generally consistent. This work has validated our central hypothesis that small-molecule inhibitors of the PPIs involving ATG5 can tune down autophagy effectively, and their pharmaceutical potential may be further explored.

Long noncoding RNA Linc00337 functions as an E2F1 co-activator and promotes cell proliferation in pancreatic ductal adenocarcinoma.

BACKGROUND: Long noncoding RNA (lncRNA) Linc00337 has been implicated in lung, gastric, colorectal and esophageal squamous cell carcinoma progression via various mechanisms; however, its clinicopathological significance and role in pancreatic ductal adenocarcinoma (PDAC) progression remains largely unknown. METHODS: Multiple approaches such as bioinformatic analysis, Transfection, quantitative real-time-PCR, Western blotting, animal studies, RNA-immunoprecipitation (RIP), RNA-pulldown and RNA-Fluorescence in situ hybridization (RNA-FISH) and were utilized to explore the role of Linc00337 in PDAC. RESULTS: Here we identified Linc00337 is an oncogenic lncRNA during PDAC progression. We found that the expression of Linc00337 is elevated in PDAC tissues and the higher Linc00337 predicts dismal prognosis. Functionally, Linc00337 promotes PDAC cell proliferation and cell cycle transition both in vitro and in vivo. Mechanistically, Linc00337 binds to E2F1 and functions as an E2F1 coactivator to trigger the targets expression during PDAC progression. CONCLUSION: Our results demonstrate a reciprocal regulation mechanism between Linc00337 and E2F1 in PDAC progression and report the clinical value of Linc00337 for PDAC prognosis and treatment.

Brca2 deficiency drives gastrointestinal tumor formation and is selectively inhibited by mitomycin C.

BRCA2 is crucial for repairing DNA double-strand breaks with high fidelity, and loss of BRCA2 increases the risks of developing breast and ovarian cancers. Herein, we show that BRCA2 is inactively mutated in 10% of gastric and 7% of colorectal adenocarcinomas, and that this inactivation is significantly correlated with microsatellite instability. Villin-driven Brca2 depletion promotes mouse gastrointestinal tumor formation when genome instability is increased. Whole-genome screening data showed that these BRCA2 monoallelic and biallelic mutant tumors were selectively inhibited by mitomycin C. Mechanistically, mitomycin C provoked double-strand breaks in cancer cells that often recruit wild-type BRCA2 for repair; the failure to repair double-strand breaks caused cell-cycle arrest at the S phase and p53-mediated cell apoptosis of BRCA2 monoallelic and biallelic mutant tumor cells. Our study unveils the role of BRCA2 loss in the development of gastrointestinal tumors and provides a potential therapeutic strategy to eliminate BRCA2 monoallelic and biallelic mutant tumors through mitomycin C.

Mitochondrial Sirtuin 3: New emerging biological function and therapeutic target.

Sirtuin 3 (SIRT3) is one of the most prominent deacetylases that can regulate acetylation levels in mitochondria, which are essential for eukaryotic life and inextricably linked to the metabolism of multiple organs. Hitherto, SIRT3 has been substantiated to be involved in almost all aspects of mitochondrial metabolism and homeostasis, protecting mitochondria from a variety of damage. Accumulating evidence has recently documented that SIRT3 is associated with many types of human diseases, including age-related diseases, cancer, heart disease and metabolic diseases, indicating that SIRT3 can be a potential therapeutic target. Here we focus on summarizing the intricate mechanisms of SIRT3 in human diseases, and recent notable advances in the field of small-molecule activators or inhibitors targeting SIRT3 as well as their potential therapeutic applications for future drug discovery.

Targeting autophagy-related protein kinases for potential therapeutic purpose.

Autophagy, defined as a scavenging process of protein aggregates and damaged organelles mediated by lysosomes, plays a significant role in the quality control of macromolecules and organelles. Since protein kinases are integral to the autophagy process, it is critically important to understand the role of kinases in autophagic regulation. At present, intervention of autophagic processes by small-molecule modulators targeting specific kinases has becoming a reasonable and prevalent strategy for treating several varieties of human disease, especially cancer. In this review, we describe the role of some autophagy-related kinase targets and kinase-mediated phosphorylation mechanisms in autophagy regulation. We also summarize the small-molecule kinase inhibitors/activators of these targets, highlighting the opportunities of these new therapeutic agents.

MicroRNA-1275 inhibits cell migration and invasion in gastric cancer by regulating vimentin and E-cadherin via JAZF1.

BACKGROUND: Emerging evidence has shown that miR-1275 plays a critical role in tumour metastasis and the progression of various types of cancer. In this study, we analysed the role and mechanism of miR-1275 in the progression and prognosis of gastric cancer (GC). METHODS: Target genes of miR-1275 were identified and verified by luciferase assay and Western blotting. The function of miR-1275 in invasion and metastasis was analysed in vitro and in vivo in nude mice. The signal pathway regulated by miR-1275 was examined by qRT-PCR, Western blotting and chromatin immunoprecipitation analyses. The expression of miR-1275and JAZF1 were measured in specimens of GC and adjacent non cancerous tissues. RESULTS: We identified JAZF1 as a direct miR-1275 target. miR-1275 supresses migration and invasion of GC cells in vitro and in vivo, which was restored by JAZF1 overexpression. Moreover, JAZF1 was recognized as a direct regulator of Vimentin. Knocking-down miR-1275 or overexpressing JAZF1 resulted in upregulation of Vimentin but downregulation of E-cadherin. Meanwhile, we validated in 120 GC patients specimens that low miR-1275expression and high JAZF1 mRNA expression levels were closely associated with lymph node metastasis and poor prognosis. The expression of JAZF1 in protein level displayed the correlations with Vimentin but inversely with E-cadherin. CONCLUSIONS: Increased miR-1275 expression inhibited GC metastasis by regulating vimentin/E-cadherin via direct suppression of JAZF1expression, suggesting that miR-1275 is a tumour-suppressor miRNA with the potential as a prognostic biomarker or therapeutic target in GC.

Knockdown of GGCT inhibits cell proliferation and induces late apoptosis in human gastric cancer.

BACKGROUND: Gamma glutamylcyclotransferase (GGCT) has been proved to be involved in various cancers, but the biological function of GGCT in gastric cancer is still largely unknown. METHODS: The expression level of GGCT was evaluated by informatics analyses based on the Oncomine database. GGCT gene was then effectively knocked down via lentivirus mediated short hairpin RNA (shRNA) system. Then a series of functional assays, including MTT, colony formation and flow cytometry analysis were conducted on gastric cancer cells following GGCT knockdown. RESULTS: We found GGCT is commonly up-regulated in gastric cancer tissues. Furthermore, MTT analysis showed that GGCT depletion significantly inhibited cell proliferation in MGC80-3 and AGS cells. Colony formation assay revealed that depletion of GGCT reduced the colony formation ability in gastric cancer cells. What's more, cell cycle analysis showed that depletion of GGCT induced gastric cancer cell cycle arrested G2/M phase. More importantly, cell apoptosis analysis further revealed that GGCT inhibition induced early and late cell apoptosis in gastric cancer. CONCLUSION: This study suggests GGCT is essential for gastric cancer proliferation and its downregulation may provide a potential anticancer therapy for gastric cancer.

Long-term outcomes after radical gastrectomy in gastric cancer patients with overt bleeding.

AIM: To investigate the difference in long-term outcomes between gastric cancer patients with and without a primary symptom of overt bleeding (OB). METHODS: Consecutive patients between January 1, 2007 and March 1, 2012 were identified retrospectively by reviewing a gastric cancer database at Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. A follow-up examination was performed on patients who underwent a radical gastrectomy. OB due to gastric cancer included hematemesis, melena or hematochezia, and gastric cancer was confirmed as the source of bleeding by endoscopy. Patients without OB were defined as cases with occult bleeding and those with other initial presentations, including epigastric pain, weakness, weight loss and obstruction. The 3-year overall survival (OS) rate, age, gender, AJCC T stage, AJCC N stage, overall AJCC stage, tumor size, histological type, macroscopic (Borrmann) type, lymphovascular invasion and R status were compared between patients with and without OB. Moreover, we carried out a subgroup analysis based on tumor location (upper, middle and lower). RESULTS: We identified 939 patients. Of these, 695 (74.0%) were hospitalized for potential radical gastrectomy and another 244 received palliative resection, rerouting of the gastrointestinal tract, chemotherapy, radiotherapy or no treatment due to the presence of unresectable tumors. Notably, there was no significant difference in the percentage of OB patients between resectable cases and unresectable cases (20.3% vs 22.1%, P = 0.541). Follow-up examination was performed on 653 patients (94%) who underwent radical gastrectomy. We found no significant difference in 3-year OS rate (68.2% vs 61.2%, P = 0.143) or clinicopathological characteristics (P > 0.05) between these patients with and without OB. Subgroup analysis based on tumor location showed that the 3-year OS rate of upper gastric cancer was significantly higher in patients with OB (84.6%) than in those without OB (48.1%, P < 0.01) and that AJCC stages I-II (56.4% vs 35.1%, P = 0.017) and T1-T2 category tumors (30.8% vs 13%, P = 0.010) were more frequent in patients with OB than in those without OB. There was no significant difference in 3-year OS rate or clinicopathological characteristics between patients with and without OB (P > 0.05) for middle or lower gastric cancer. CONCLUSION: Upper gastric cancer patients with OB exhibited tumors at less advanced pathological stages and had a better prognosis than upper gastric cancer patients without OB.

Expression of Fatty Acid Synthase Negatively Correlates with PTEN and Predicts Peritoneal Dissemination of Human Gastric Cancer.

BACKGROUND: This study aimed to examine the clinical significance of fatty acid synthase (FASN) expression in gastric cancer (GC), and investigate any prognostic role. MATERIALS AND METHODS: FASN expression was assessed in gastric cancers by immunohistochemistry using 60 paraffin-embedded tissue specimens, and clinical data were collected by retrospective chart review. Moreover, FASN mRNA expression in 15 fresh resected specimens was evaluated by the reverse transcription-polymerase chain reaction (RT-PCR). Immunohistochemical staining of PTEN was performed to assess the correlation of PTEN with FASN in gastric cancer. RESULTS: Increased expression of FASN was noted in gastric cancers. The frequency of FASN gene amplification was also significantly higher in gastric cancer than in adjacent normal tissue. FASN expression in human gastric cancer tissues was significantly correlated with patient TNM stage and peritoneal dissemination (p<0.05). Moreover, higher FASN expression significantly correlated with shorter overall survival (p<0.05). Here, upregulation of FASN negatively correlated with PTEN expression in gastric cancer. CONCLUSIONS: These findings indicate that FASN expression is upregulated in gastric cancer, and increased FASN may be critical to th peritoneal metastasis and survival. Our results suggest that FASN upregulation and PTEN downregualtion may be involved in peritoneal dissemination for gastric cancer progression.

Loss of PCDH9 is associated with the differentiation of tumor cells and metastasis and predicts poor survival in gastric cancer.

Microarray studies revealed down-regulation of PCDH9 mRNA level in lymph node metastasis of gastric cancer compared with the primary tumors. The expression of PCDH9 protein and its clinicopathological relevance were assessed on tissue microarrays of 1072 cases of gastric cancer. Its prognostic value was further evaluated on a small cohort of 175 gastric cancers. PCDH9 was down-regulated during the development and progression of gastric cancer. The overall rates of PCDH9 expression in normal, primary tumor, nodal and hepatic metastatic tissues were 100 % (1072/1072), 48.0 % (515/1072), 20.1 % (34/169), and 5.6 % (1/18), respectively. Positive staining of PCDH9 protein was significantly reversely correlated with tumor size, tumor differentiation, tumor invasion, lymph node metastasis, and disease progression. The Cox proportional hazards model revealed that the PCDH9 was an independent prognostic factor for gastric cancer. Therefore, decreased expression of PCDH9 is frequent in human gastric cancer metastasis and PCDH9 expression is an independent prognostic factor, suggesting that PCDH9 could be a promising biomarker of this malignancy.

Cyclooxygenase-2 inhibition as a strategy for treating gastric adenocarcinoma.

Cyclooxygenase-2 (COX-2) has been proven to play critical roles in inflammation as well as in cancer. Some studies have shown that the anti-inflammatory, immunosuppressive and anti-arthritic effects of celecoxib are mainly attributed to the inhibition of COX-2 expression. The present study aimed to investigate the function of COX-2 in human gastric adenocarcinoma (GAC). Forty-five cases of human GAC tissues and corresponding adjacent non-cancerous tissues (ANCTs) were collected. The expression of COX-2 and proliferating cell nuclear antigen (PCNA) was assessed using immunohistochemical assay through a tissue microarray procedure. GAC cells (SGC-7901 and MKN-45) in vitro were treated with COX-2 siRNA or different concentrations of celecoxib to observe their effects on cell proliferation, invasion and the underlying molecular mechanisms. As a consequence, the expression of COX-2 and PCNA was found in cancer tissues with a higher strong reactivity rate, compared with the ANCTs (80.0 vs. 53.3%, P=0.011; 68.9 vs. 48.9%, P=0.047), and COX-2 was positively associated with lymph node metastasis of GAC patients (P=0.011). Targeted knockdown of COX-2 inhibited the proliferation, migration and invasion of GAC cells with decreased expression of PCNA. COX-2 inhibitor celecoxib also suppressed the proliferative activities of GAC cells with decreased expression of COX-2 and PCNA. In addition, the tumor volume in the MKN-45 subcutaneous tumor model treated with siCOX-2 was significantly smaller than that of the negative control (NC) group (P<0.01). Taken together, our findings offer a strong preclinical rationale to target COX-2 signaling as a therapeutic strategy to improve the treatment of gastric adenocarcinoma.

CD86 gene variants and susceptibility to pancreatic cancer.

BACKGROUND: Pancreatic cancer is one of the most lethal cancers worldwide. CD86 (B7-2) is a costimulatory molecule on antigen-presenting cells and plays critical roles in tumor immunity. It has been reported that polymorphisms in CD86 gene can be involved in the development of various cancers. Here, we investigated the association of two CD86 polymorphisms, +1057G/A (rs1129055) and +2379G/C (rs17281995), with pancreatic cancer in the Chinese population. METHODS: The two polymorphisms were identified in 369 pancreatic cancer patients and 412 healthy controls using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Data were analyzed by chi-square test and adjusted for body mass index, smoking, drinking, and diabetes status. RESULTS: Results showed that the frequency of the +1057A allele was significantly higher in pancreatic cancer cases than in controls (59.8 vs. 52.8 %, p = 0.021). Comparison of genotype frequencies showed that +1057GA and +1057AA genotypes were significantly increased in the pancreatic cancer group (odds ratio (OR) = 1.52; 95 % confidence interval (CI), 1.09-2.38; p = 0.026; and OR = 1.90; 95 % CI, 1.21-3.01; p = 0.007). We did not find any association between the +2379G/C polymorphism and pancreatic cancer. Analysis of haplotypes indicated that the AG (+1057, +2379) haplotype was correlated with the susceptibility to this disease (p = 0.019). CONCLUSIONS: These results suggest that the CD86 +1057G/A polymorphism and AG (+1057, +2379) haplotype are genetic risk factors for pancreatic cancer.

Expression of phosphorylated mammalian target of rapamycin in colorectal cancer and its significance / 第二军医大学学报

Objective: To explore the roles of mammalian target of rapamycin (mTOR) and the activated mTOR (phosphorylated mTOR, p-mTOR) in the development and progression of colorectal cancer, and to discuss the clinical significance. Methods: The expression of mTOR and p-mTOR in 185 coLorectal cancer specimens and the corresponding adjacent tissues were evaluated by tissue microarray and immunohistochemistry,and the relationship between the expression and the age, sex, invasion depth( T stage) ,lymph metastasis, TNM stage and differentiation degree was analyzed. Results1 Diffused expression of mTOR and hardly any expression of p-mTOR were found in the adjacent tissues. The expression of mTOR and p-mTOR was obviously stronger in the colorectal cancer tissues compared with that in the adjacent tissues. The over-expression rates of mTOR and p-mTOR in colorectal cancer were 45. 9% and 42. 2%, respectively. There was no significant correlation of mTOR and p-mTOR over-expression with age, sex( P> 0. 05); the over-expression of mTOR was correlated with the differentiation degree (P0. 05). The correlation of p-mTOR over-expression with the invasion depth (T stage) ,lymph metastasis and TNM stage was significant (P0. 05). Conclusion:Over-expression of p-mTOR is closely associated with the malignant phenotype of colorectal cancer. It is also indicated that p-mTOR may be involved in the development and progression of colorectal cancer.