Metadherin inhibits chemosensitivity of triple-negative breast cancer to paclitaxel via activation of AKT/GSK-3ß signaling pathway.

Triple-negative breast cancer (TNBC) has an aggressive clinical course, and paclitaxel (PTX)-based chemotherapy remains the main therapeutic drug. Metadherin (MTDH) acts as an oncogene that regulates proliferation, invasion, metastasis, and chemoresistance. This study aimed to investigate whether TNBC chemosensitivity to PTX was related to the MTDH/AKT/glycogen synthase kinase-3beta (GSK-3ß) pathway. Clinical baseline characteristics and immunohistochemistry (IHC) were used to evaluate the expression and prognosis of MTDH and AKT (protein kinase B, PKB) in TNBC patient samples. MTDH shRNA, MTDH overexpression vector, MK-2206, and PTX intervention were used in cell models and mouse tumor-bearing models. Afterwards, mRNA and protein levels were assessed using quantitative real-time polymerase chain reaction and Western blot. Evaluate the level of tumor cell apoptosis and cell cycle using flow cytometry. Cell viability was detected using Cell Count Kit 8. The in vivo imaging system is used to analyze the growth of tumors. We found that higher expression of MTDH or AKT resulted in poorer disease-free survival and a lower Miller-Payne grade. MTDH promotes cell proliferation and increases p-AKT and p-GSK-3ß expression in TNBC cells. Notably, suppression of AKT terminated MTDH overexpression-induced cell proliferation and apoptosis. MTDH knockdown or the AKT inhibitor MK2206 reduced the p-AKT and p-GSK-3ß ratio, reduced cell viability and proliferation, increased cell apoptosis, and increased chemosensitivity to PTX. In vivo, xenograft tumors of an MTDH knockdown+MK2206 group treated with PTX were the smallest compared to other groups. In short, MTDH inhibits TNBC chemosensitivity to PTX by activating the AKT/GSK-3ß signaling pathway.

Calcium dobesilate prevents PLD-induced hand-foot syndrome by alleviating capillary endothelial tight junction injury via the HA/CD44 pathway.

Pegylated liposomal doxorubicin (PLD) has excellent therapeutic efficacy in the treatment of cancers, but can cause serious adverse reactions such as hand-foot syndrome (HFS). Our previous research suggests that both PLD-induced HFS may be associated with injury to tight junctions (TJs) in the skin and that calcium dobesilate (CaD) can alleviate HFS. However, the underlying molecular mechanism is not well understood. Here, we created an in vitro PLD-treated model using Human Microvascular Endothelial Cell line-1 (HMEC-1) and an in vivo HFS rat model to investigate the underlying pathways. Treatment with PLD increased the expression of HYAL-1, CD44, and hyaluronic acid (HA) concentration, while reducing ZO-1 and Claudin-5 expression. Moreover, PLD treatment induced the degradation of higher molecular weight HA to its lower molecular weight counterpart, elevating the permeability of both HEMC-1 cell membranes and rat paw skin capillaries. AD-01 (CD44 inhibitor) inhibited the effect of PLD on the expression of ZO-1 and Claudin-5. Furthermore, CaD treatment suppressed the expression of HYAL-1 and CD44, mitigated HA degradation, and enhanced the expression of ZO-1 and Claudin-5. This resulted in decreased permeability in HEMC-1 cells and rat skin capillaries. In summary, our data suggest that PLD may promote the destruction of TJs via the HA/CD44 pathway, thereby leading to HFS through increased skin permeability and exacerbated doxorubicin extravasation. Moreover, CaD can inhibit this pathway, offering a potential therapeutic avenue to alleviate HFS.

Body Mass Index at Diagnosis as a Prognostic Factor for Early-Stage Invasive Breast Cancer after Surgical Resection.

BACKGROUND: The aim of this study was to investigate the association between body mass index (BMI) and prognosis of Chinese women with breast cancer. PATIENTS AND METHODS: 3,380 primary breast cancer patients who underwent surgery from 2010 to 2012 were selected and classified as low BMI group (BMI < 25.0) and high BMI group (BMI ≥ 25.0). The follow-up data for disease-free survival (DFS) and overall survival (OS) were obtained from 3,178 patients (median follow-up of 58 months). Cox regression analysis was used to evaluate the effect of BMI on DFS and OS. RESULTS: The high BMI group showed more aggressive pathological features. BMI was negatively associated with OS (hazard ratio (HR) 1.33, 95% confidence interval (CI) 1.06-1.66; p = 0.012) but not DFS (HR 1.15, 95% CI 0.94-1.40; p = 0.17). Furthermore, when stratified by age, BMI was significantly and negatively associated with OS (HR 1.43, 95% CI 1.05-1.95; p = 0.025) in patients above 50 years of age, but this effect was not detected in younger patients. CONCLUSION: BMI was an independent prognostic factor of OS in Chinese women with breast cancer, and age might be a mitigating factor. Among patients above 50 years of age, those with a high BMI were at greater risk of poor prognosis compared to individuals with a low BMI.

[Proteomic identification and comparison of differentiation-related proteins in gastric carcinoma cell lines].

OBJECTIVE: To investigate the differentiation-related proteins in human gastric carcinoma cell lines by comparative proteomics. METHODS: The holoproteins of human gastric carcinoma cell lines MKN28, SGC7901 and BGC823 were measured by two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Some proteins identified by proteomics were tested by Western blot in the cell strains and tissues of gastric carcinoma. RESULTS: 14 differential protein spots were found in the 3 gastric carcinoma cell lines, among them 8 spots were identified by MALDI-TOF-MS. These proteins were probably thioredoxin peroxidase, glyceraldehyde-3-phosphate dehydrogenase (GAPD), beta-tubulin polypeptide, hypothetical protein, zinc finger protein (ZNF) 139, protein-tyrosine kinase, calreticulin precursor, and tropomyosin, proteins related with biological behavior of gastric carcinoma cells such as signal transduction, cellular homeostasis, glycolysis, antioxidation action, multidrug resistance(MDR), etc. The expressions of those proteins in gastric cancer cells and tissues identified by Western blot were consistent with the results obtained by proteomics. CONCLUSION: Differential proteins are found in 3 human gastric carcinoma cell lines, mainly, proteins related with cell signaling, maintenance of homeostasis, glycolysis, metabolism of anti-cancer drug and anti-oxidative injury, etc.

[Relationship of cyclooxygenase-2 and multidrug resistance associated factors to chemosensitivities in gastrointestinal carcinomas].

OBJECTIVE: To investigate the relationship of cyclooxygenase-2 (COX-2), p-glycoprotein(P-gp), glutathione S-transferase-pi (GST-pi), and topoisomerase II alpha (Topo II alpha) to chemosensitivities in gastrointestinal tract carcinomas. METHODS: The tumor tissue samples were collected from 84 specimens of gastrointestinal carcinomas. The expressions of COX-2, P-gp, GST-pi, and Topo II alpha were determined immunohistochemically. The chemosenisitivity of each sample to 9 drugs were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RESULTS: The positive rates of COX-2, P-gp, GST-pi and Topo II alpha were 48.8%, 76.2%, 78.6% and 66.7% respectively. The expression of COX-2 was correlated with the expression of P-gp and Topo II alpha significantly (r = 0.287, 0.256, both P < 0.05). In terms of relationships of four MDR-related factors expression to inhibition rate of tumor cells, the inhibition rates of PTX, eADM and OPT in COX-2 strong expression group were significantly lower than those in COX-2 weak expression group (t = 2.21, 3.11, 2.09; all P < 0.05). The inhibition rates of PTX, OXA and DDP in P-gp strong expression group were lower than those in weak group (t = 2.54, 2.47, 2.05; all P < 0.05). There were lower inhibition rates for 5-Fu in GST-pi strong expression group (t = 2.13, P < 0.05), and higher inhibition rates for VCR, PTX and eADM in TopoII alpha strong expression group (t = -2.29, -2.12, - 2.26, all P < 0.05). CONCLUSIONS: The overexpressions of MDR-related factors in gastrointestinal carcinomas were only associated with the chemosensitivity to some of the chemotherapeutic agents, but not all. MDR related factors may be not specific and accurate predictors for the clinical chemosensitivity.

[Effect of ulinastatin on intestinal mucosal barrier function of rats with obstructive jaundice].

OBJECTIVE: To investigate the effect of ulinastatin on intestinal mucosal barrier function of rats with obstructive jaundice. METHODS: Seventy-two male SD rats were randomly divided into sham operation, obstructive jaundice, and ulinastatin treatment groups (groups A, B, and C, respectively). In groups B and C, the common bile duct was ligated to induce obstructive jaundice. The rats in group C were given intraperitoneal injection of ulinastatin at the daily dose of 40,000 IU/kg after the operation, while those in groups A and group B received equal amount of normal saline. At 3, 5, 7 and 10 days after the operation, the liver function and plasma endotoxin level were evaluated and measured, and bacterial culture of the mesenteric lymph nodes, liver and spleen was performed. The terminal ileum mucosa was observed under light microscope, and the intestinal villi and mucosal thinckness was examined with image analysis system. RESULTS: The indices relative to the liver function and plasma endotoxin level were higher at different time points of observation in group B than in group A (P<0.01), and were lower in group C than in group B (P<0.01). Plasma endotoxin level was similar between groups A and C 3 days after the operation (P>0.05). The rate of bacterial translocation was higher in group B than in group A and C (P<0.01, P<0.05), but comparable between groups A and C (P>0.05). Intestinal mucosal injury was observed in group B 3 days after operation, and aggravated with the passage of time. The injury was milder in group C. The intestinal villus length and mucosal thickness were greater in groups A and C than in group B (P<0.01 or P<0.05), but comparable between the former two groups 3 days after operation (P>0.05). CONCLUSION: In early stage of obstructive jaundice, the intestinal mucosal barrier may sustain injuries which aggravate with time; ulinastatin has significant effect in protecting the mucosal barrier function especially against early pathological changes.

[Detecting bone marrow micrometastasis of gastric cancer by magnetic activated cell sorting combined with fluorescent activated cell sorting].

BACKGROUND & OBJECTIVE: Several methods are used to detect bone marrow micrometastasis of gastric cancer with different accuracies. In breast cancer, tumor cells in blood can be detected sensitively and specifically by magnetic activated cell sorting (MACS) and fluorescent activated cell sorting (FACS). This study was to investigate the clinical value of this method in detecting bone marrow micrometastasis of gastric cancer. METHODS: Thirty-five patients, who received operation for gastric cancer from Dec. 2002 to Jun. 2003, were selected. Mononuclear cells were separated from their bone marrows. After marked by MACS minibeads conjugated with cytokeratin (CK) 7/8 antibodies, anti-CK-fluorescein isothiocyanate (FITC), and anti-CD45-perdinin chlorophyll protein (PerCP), tumor cells were enriched twice by MS+/RS+ positive separation column. FACS analysis was conducted on these samples before and after MACS enrichment. The results were compared with clinicopathologic parameters. RESULTS: Disseminated tumor cells were detected in bone marrow of 3 samples(8.6%) before MACS enrichment, and 25 samples (71.4%) after enrichment. The frequencies of tumor cells were 1.4 x 10(-8)-2.4 x 10(-5), 2.2 x 10(-7) -3.7 x 10(-5), and 4.0 x 10-(6)-8.6 x 10(-5) in patients with moderately differentiated, poorly differentiated, and undifferentiated carcinoma, respectively, with significant differences (P = 0.026). Bone marrow micrometastasis positively correlated with tumor TNM stage (P = 0.008), while had no correlation with tumor size, depth of wall invasion, and other clinicopathologic parameters. CONCLUSIONS: MACS combined with FACS may improve detection rate of bone marrow micrometastasis of gastric cancer. The patients with poor differentiation and in advanced TNM stage have more disseminated tumor cells in bone marrow.

[Multi-antibody combined determination of lymph node micrometastasis in patients with gastric cancer].

BACKGROUND & OBJECTIVE: Immunohistochemical staining is a simple method for determination of lymph node micrometastasis in gastric cancer; but the sensitivity is low. Whether this disadvantage can be improved using multi-antibody combined determination is still controversial. This study was designed to determine the lymph node micrometastasis in patients with gastric cancer by the antibodies of cytokeratin 20 (CK20), epithelial membrane antigen (EMA), and carcinoembryonic antigen 72-4 (CA72-4)for clarifying the value of multi-antibody combined determination of micrometastasis. METHODS: A total of 466 lymph nodes was collected with operation from 44 gastric cancer patients from April 1991 to April 1994. All these lymph nodes showed no lymph node metastasis by routine histological examination. Immunohistochemical staining was performed on all the samples by the mouse antibodies of anti-CK 20, mouse anti-EMA, and mouse anti CA72-4, respectively. Then, the micrometastases were identified under microscope according to the color of the cells. The results were analyzed according to clinical, pathological, and follow-up data. RESULTS: Fifty-one (10.9%) lymph nodes of 18 (40.9%) cases showed micrometastasis. The number of micrometastatic lymph nodes were detected by CK20, CA72-4, and EMA was 40 (8.6%), 27 (5.8%), and 21 (4.5%), respectively. There was no significant correlation between the lymph node micrometastasis and clinical data, such as gender, age, tumor site, histological differentiation, and stage (P >0.05). The 5-year survival rate of the patients with lymph node micrometastasis was lower than that of the patients without lymph node micrometastasis(61.11+/-11.49% vs. 92.31+/-5.23%, P=0.0113). The 5-year survival rate of the patients with 3 or more positive micrometastatic nodes was lower than that of the patients with less than 3 positive micrometastatic nodes (44.44+/-16.56% vs. 77.78+/-13.86%,P=0.0196). CONCLUSION: Immunohistochemical staining marked by different antibodies can be an useful method to seek more micrometastatic lymph nodes in gastric cancer patients.

[Drug distribution in gastric cancer and adjacent tissues by preoperative intraperitoneal chemotherapy with Co-fluorouracil liposome].

OBJECTIVE: To examine the distribution of fluorouracil in gastric cancer (CA), lymph node (LN), normal gastric mucosa (NG), peritoneum (PE), greater omentum (GO) and lesser omentum (LO) by preoperative intraperitoneal chemotherapy with Co-fluorouracil liposome (Co 5-Fu), and offer an experimental basis for clinic practice. METHODS: Ninety-six gastric cancer patients were divided into four groups: Co 5-Fu i.v. injection group (Co 5-Fu i.v.), Co 5-Fu intraperitoneal perfusion group (Co 5-Fu i.p.), 5-Fu i.v. injection group (5-Fu i.v.) and intraperitoneal perfusion group (5-Fu i.p.) given on day-2, day-1 and 60 minutes before operation. Fluorouracil concentration in all tissues collected during operation were examined by high performance liquid chromatography (HPLC). RESULTS: The fluorouracil concentration in the tissues in Co 5-Fu i.p. group was significantly higher than that in Co 5-Fu i.v. or 5-Fu i.p. group (P < 0.05 or P < 0.01), and that in 5-Fu i.p. group was greatly higher than that at 5-Fu i.v. group (P < 0.01). In Co 5-Fu i.p. group, the concentration of drug in LN, CA, PE, NG, GO and LO decreased gradually with the former 3 tissues significantly higher than the latter 3 tissues (P < 0.01), and adjacent lymph node was the highest. In Co 5-Fu i.v. group, the ranking was LN, CA, NG, PE, GO and LO with the former 3 tissues significantly higher than the latter 3 tissues (P < 0.01) and showing tumor tissues higher than the other tissues (P < 0.01). In 5-Fu i.p. group, the ranking was PE, LN, CA, NG, GO and LO with the former 2 tissues significantly higher than the latter tissues (P < 0.01). CONCLUSION: Co 5-Fu possesses drug targeting, slow release and long effect in gastric cancer tissues and adjacent lymph nodes. Preoperative chemotherapy with Co 5-Fu i.p. is more advantageous than 5-Fu given i.v. or 5-Fu i.p.

[Relationship between expression of multiple tumor suppressor(MTS1) and E-cadherin and metastasis of breast carcinoma].

BACKGROUND & OBJECTIVE: Most of breast cancer patients died of distant metastasis. There is still not a method of predicting and early diagnosing the metastasis. This study was conducted to investigate the relationship between the expression of multiple tumor suppressor 1(MTS1) as well as E-cadherin and metastasis in breast cancer. METHODS: The expression of MTS1 and E-cadherin were detected by SP immunohistochemical technique in 54 paraffin-embedded tissue specimens of breast cancer. RESULTS: The positive rates of MTS1 in the distant metastasis group and more than 4 local lymph node metastasis group were 40.7% and 38.1%, respectively, which were lower than individual corresponding control groups (74.1% and 69.7%) (P< 0.05). The MTS1 expression rates in the three groups of pathology grade I, II, and III were 72.6%, 58.8%, and 31.3%, respectively, with significant differences (P< 0.05). The positive rates of E-cadherin in the distant metastasis group and more than 4 local lymph node metastasis group were 37% and 23.8%, respectively, which are lower than individual corresponding control groups (70.4% and 66.7%) (P< 0.05). The E-cadherin expression rates in the three groups of pathology grade I, II, III were 80.9%, 47.1%,and 25%, respectively, with significant differences (P< 0.01). CONCLUSION: There was a close relation between MTS1, E-cadherin and invasion, metastasis of breast cancer. They can be used as the biological markers for evaluating the latent metastasis, disease development, and prognosis in breast cancer.