Autophagic regulation of cell growth by altered expression of Beclin 1 in triple-negative breast cancer.

Beclin 1 is a promoter gene for autophagy as well as a key factor for regulating tumor cell growth and death. Allelic deletion of Beclin 1 has been observed in certain triple-negative breat cancer (TNBC) cells, and it might be associated with increased proliferation and invasion in TNBC cells. In this study we investigated the relationship between Beclin 1 expression and prognosis for TNBC patients, as well as the influence on cell growth by Beclin 1 overexpression in different cultural conditions. Beclin 1 expression in TNBC tissues was measured by immunohistochemical staining and correlated with clinicopathologic parameters for TNBC patients. The plasmid of pDS-RED-C1-Beclin 1 was transfected to BT-549 and MDA-MB-231 cells and autophagy, proliferation, apoptosis, cell cycle and Epithelial-mesenchymal transition (EMT) process were measured. Results indicated that high level of Beclin 1 expression was correlated with more lymph nodes and distant metastasis but unrelated to survival rates in 5 years for TNBC patients. In vitro, overexpression of Beclin 1 improved cellular autophagy in both BT-549 and MDA-MB-231 cells, inhibited cell proliferation at normal cultural condition and increased cell survival in starvation, hypoxia or with doxorubicin stimulation. Besides, Beclin 1 overexpression decreased cell apoptosis, induced cells to be in G0/G1 phase and promoted EMT process through Wnt/ß-catenin pathway in starvation. Thus, Beclin 1 overexpression plays a double role in BT-549 and MDA-MB-231 cell growth by elevating the capability of autophagy. These findings might be useful for searching a proper method for clinical therapy of TNBC from the aspect of autophagy in future.

Preliminary Study on the Expression and the Clinical Significance of CD133 in Peripheral Blood of Patients with Gastric Adenocarcinoma.

Background. Significances of CD133 mRNA in peripheral blood mononuclear cells (PBMCs) of gastric adenocarcinoma (GC) patients were investigated. Methods. Correlations of CD133 mRNA expression in PBMCs on clinicopathological parameters or CD133 protein expression were analyzed. Receiver operating characteristic curve according to bright scale value (BSV) of CD133 mRNA was used to group patients for prognosis analysis. Results. BSV of preoperative CD133 mRNA in PBMCs in GC was significantly higher than that in volunteers or in GU. Invasive depth or metastatic lymph node number for higher BSV of preoperative CD133 mRNA and invasive depth or lymphatic vessel invasion for higher BSV of postoperative CD133 mRNA in the PBMCs were identified. Patients with CD133(+) expression in primary lesion had a significantly higher expression of preoperative CD133 mRNA in the PBMCs. The expression of preoperative or postoperative CD133 mRNA in PBMCs related positively to CD133 mRNA expression in primary lesion. Patients with higher expression of preoperative or postoperative CD133 mRNA shared significantly shorter survival compared with that in lower expression group. Conclusion. Higher levels of preoperative or postoperative CD133 mRNA in PBMCs of GC correlated positively to the lymphatic metastasis and the BSV of CD133 mRNA in primary lesion, indicating the poorer survival.

Expressions and clinical significances of c-MET, p-MET and E2f-1 in human gastric carcinoma.

BACKGROUND: To investigate on the expressions and the clinical significances of hepatocyte growth factor receptor (c-MET), phosphorylated c-MET (p-MET) and e2f-1 transcription factor in primary lesion of gastric adenocarcinoma (GC). METHOD: Tissue samples from the primary lesion of GC in patients who accepted D2/D3 radical gastrectomy with R0/R1 resection were stained by immunohistochemistry of c-MET, p-MET, e2f-1 and Ki-67. The univariate and the multivariate analyses involving in clinicopathological parameters and prognostic factors were evaluated. RESULTS: The positivity rates for c-MET (66.12%, 80 cases/121 cases), p-MET (59.50%, 72 cases/121 cases), e2f-1 (38.84%, 47 cases/121 cases) and Ki-67 (72.73%, 88 cases/121 cases) in primary lesion of GC was significantly higher than that in non-cancerous tissue at 5 cm places far from the margin of primary lesion (P < 0.05, respectively). The deeper tumor invasion, the severer lymph node metastasis, the later stage of TNM and the higher expression of Ki-67 was respectively an independent risk factor for the higher expression of c-MET or p-MET, but the younger age and the shorter survival time was an independent risk factor for the higher expression of e2f-1 respectively. Survival analysis showed that the worse prognosis could be observed in the patients with the combination of both c-MET-positive and e2f-1-negative (P = 0.038) or both p-MET-positive and e2f-1-negative (P = 0.042). Cox analysis demonstrated that the severer lymphatic node metastasis and the higher positivity rate of c-MET, p-MET or e2f-1 were an independent prognosis factor respectively. The higher expression of e2f-1 was identified in patients with Stage I-II, which correlated with a shorter survival time. Survival analysis also revealed that the prognosis of patients with positive expression of e2f-1 at Stage I-II was significantly worse than that in patients with negative expression of e2f-1 (χ2 = 13.437, P = 0.001). However, in the cases with Stage III-IV, no significant difference could be identified in the prognostic comparison between positive and negative expressions of e2f-1. CONCLUSIONS: The expression of c-MET or p-MET is an independent prognosis factor. It has been observed that the higher expression of e2f-1 occurred in the early stages while the lower expression of it in the later stages in GC.

Expression and significances of contactin-1 in human gastric cancer.

Background. This study aimed at determining the relationship between vascular endothelial growth factor-C (VEGF-C), vascular endothelial growth factor receptor-3 (VEGFR-3), and contactin-1 (CNTN-1) expression in gastric cancer (GC). Methods. The expression level of CNTN-1 mRNA and CNTN-1 protein of 33 cases was determined using RT-PCR and Western Blot. And 105 cases were immunohistochemically examined for VEGF-C, VEGFR-3, and CNTN-1 expressions. Assessment of lymphatic vessel density (LVD) was also performed by D2-40 immunostaining. Then we analyzed the relationships between VEGF-C, VEGFR-3, and CNTN-1, as well as their correlations with clinicopathologic features, LVD, and survival time. Results. The positivity rate of VEGF-C, VEGFR-3, and CNTN-1 in primary tumor was 56.19%, 64.76%, and 58.09%. The expression of CNTN-1 significantly correlated with VEGF-C (P < 0.001) and VEGFR-3 (P < 0.001). All of them were closely related to TNM stage, lymphatic invasion, and lymph node involvement (P < 0.05). LVD was significantly correlated with VEGF-C (P = 0.001), VEGFR-3 (P = 0.011), and CNTN-1 expression (P < 0.001). VEGF-C, VEGFR-3, and CNTN-1 expression significantly associated with poorer prognosis (P < 0.001, P = 0.034, P = 0.012, resp.). Conclusion. CNTN-1 associated with VEGF-C and VEGFR-3 expression in GC. All of them correlated with lymphatic metastasis, which might play an important role in the lymphatic invasion via lymphangiogenesis pathway in GC.

Efficacy and safety of linezolid in the treatment of extensively drug-resistant tuberculosis.

Linezolid is a new antibiotic with activity against Mycobacterium tuberculosis in vitro and in vivo. This study aims to evaluate the efficacy and safety of linezolid in the treatment of extensively drug-resistant tuberculosis (XDR-TB). We used a linezolid-containing regimen in the treatment of 14 XDR-TB patients. Two years of individualized chemotherapy regimens were adopted on the basis of the patients'medication history and the results of drug susceptibility testing. The patients received 600 mg of linezolid twice a day for the first 1-2 months, followed by once a day thereafter. Eleven patients (78.6%) showed significant improvement in clinical symptoms. Chest computed tomography revealed that 10 patients (71.4%) showed cavity closure. Smear conversion and culture conversion were achieved in all 14 patients (100%) with an average of 64 and 63 days, respectively. The exact proportions of serious and minor adverse events determined by linezolid were 21.4% (3/14) and 64.3% (9/14), respectively. These data show that linezolid-containing chemotherapy for the treatment of XDR-TB may significantly improve clinical symptoms, promote lesion absorption and cavity closure, and accelerate sputum conversion. Further, adverse reactions can be tolerated and resolved with suitable intervention.

Study on lymph node metastasis correlated to lymphangiogenesis, lymphatic vessel invasion, and lymph node micrometastasis in gastric cancer.

BACKGROUND: This study aimed to evaluate the clinical significance of lymphangiogenesis, lymph vessel invasion (LVI), and lymph node (LN) micrometastasis (LNMM) in patients with gastric cancer. METHODS: The influences of the expression levels of LVI, lymph vessel density (LVD) by D2-40 immunohistochemical (IHC) staining (n=68), LNMM (including CK 20 and CK pan immunostainings, n=51) on the clinicopathologic profiles and the prognosis were analyzed. RESULTS: The higher positive rate of LVI-IHC was related to deeper invasion (P=0.044), later TNM stage (P=0.003), and more extensive LN metastasis (LNM, P=0.000). The level of LVD was significantly associated with venous invasion (P=0.037), later TNM stage (P=0.020), positive LVI-HE (P=0.040), positive LVI-IHC status (P=0.001), and severer LNM (P=0.001). Better prognosis in LVI negative group than LVI positive group has been identified. The survival rate of the group with LVD≥15/field was significantly lower than that in the group with LVD≤14/field (P=0.032). Invasion depth, N stage, LNM, blood vessel invasion, or LVI was respectively an independent prognostic factor to 3-y survival rate. The incidence of patients with LNM and metastasized LNs increased respectively from 74.5% (38/51) by HE staining to 88.2% (45/51) by CK immunostaining and from 32.0% (253/791) to 41.5% (328/791) (P=0.001). The increment of LNMM was correlated to larger tumor diameter (P=0.001), deeper invasion (P=0.018), LNM (P=0.001) and later TNM stage (P=0.012), positive LVI (P=0.04). Meanwhile, the evaluation on LNMM revealed the migration of LN stage (N(0)→N(1) in seven patients, N(1)→N(2) in six patients, and N(2)→N(3) in one patient), and TNM stage (I(b)→II in four patients, II→III(a) in 4 patients, III(a)→III(b) in 3 patients, and III(b)→IV in one patient). Survival analysis demonstrated that better prognosis in patients without LNM and/or LNMM. CONCLUSION: Our immunohistochemical analyses using antibodies of D2-40 and CK, including both CK 20 and CK pan, detected a higher incidence of LVIs and LNMs in gastric cancer specimens. This study shows close correlations among lymphangiogenesis related factors, such as LVI, LVD, and LNMM, and patients' prognosis after surgery. Therefore, immunohistochemical evaluations of these factors could be used for the accurate determination of tumor aggressiveness.

Influencing factors and clinical significance of the metastatic lymph nodes ratio in gastric adenocarcinoma.

BACKGROUND: To investigate influencing factors of the metastatic lymph nodes ratio (MLR) and whether it is related to survival in patients with gastric adenocarcinoma. METHODS: We retrospectively evaluated the clinical features of 121 patients with gastric adenocarcinoma enrolled in our hospital between 2000 and 2007. The receiver operating characteristic (ROC) curve was used to determine the cutoff of the MLR, and CK20 immunohistochemical staining was used to detect micrometastasis of the lymph nodes. RESULTS: The areas under the ROC curve of MLR used to predict the death of 3-year and 5-year postoperative patients were 0.826 +/- 0.053 and 0.896 +/- 0.046. Thus MLR = 30.95% and MLR = 3.15% were designated as cutoffs. The MLR was then classified into three groups: MLR1 (MLR<3.15%); MLR2(3.15% < or = MLR < or = 30.95%); and MLR3 (MLR>30.95%). We found that patients with a higher MLR demonstrated a much poorer survival period after radical operation than those patients with a lower MLR (P = 0.000). The COX model showed that MLR was an independent prognostic factor (P = 0.000). The MLR could also discriminate between subsets of patients with different 5-year survival periods within the same N stage (P < 0.05). The MLR has been shown to be 34.7% (242/697) by HE staining and 43.5% (303/697) by CK staining (P = 0.001). The clinicopathological characteristics of lymph vessel invasion and the depth of invasion could significantly affect the MLR. CONCLUSION: MLR is an independent prognostic factor in gastric cancer. The combined ROC curve with MLR is an effective strategy to produce a curve to predict the 3-year and 5-year survival rates.