Loss of Atg7 in Endothelial Cells Enhanced Cutaneous Wound Healing in a Mouse Model.

BACKGROUND: Emerging evidence has linked autophagy to skin wound healing; however, the underlying cellular and molecular mechanisms remain poorly understood. The present study was designed to determine the role of autophagy in endothelial cell (EC)-mediated skin wound healing in mice. METHODS: Autophagy-related gene (Atg7) in mouse ECs was inactivated by the Cre-loxP system under the control of an EC-specific VE-Cadherin (Cdh5) promoter (Atg7EC-/- mice). Full-thickness skin wounds were created on the dorsum of wild-type (WT), Cdh5-Cre+, floxed Atg7 (Atg7F/F), and Atg7EC-/- mice. Autophagic activity was determined by autophagic flux assay in the primary culture of ECs isolated from these mice. The wound re-epithelialization and angiogenesis was examined by histological analyses. The angiogenic activity of ECs was evaluated by tube formation assay in vitro. EC proliferation was examined by a cell count CCK-8 kit. EC-originated intercellular communication with dermal fibroblasts and keratinocytes was assessed by measuring the effect of EC conditional medium on the growth of keratinocytes and fibroblasts. The levels of VEGF, EGF, bFGF in EC conditional medium were measured by ELISA. RESULTS: Autophagy deficiency in ECs markedly enhanced the re-epithelialization and the wound closure during skin wound healing. However, it has minimal impact on angiogenesis in the wounded skin. Notably, autophagy deficiency in ECs did not affect their proliferation and migration or angiogenic activity per se but enhanced the EC conditional medium-induced proliferation and migration of keratinocytes and fibroblasts. CONCLUSIONS: These results demonstrate for the first time an inhibitory role of autophagy in the EC-originated paracrine regulation of skin wound healing.

HOXA10 promotes cell invasion and MMP-3 expression via TGFß2-mediated activation of the p38 MAPK pathway in pancreatic cancer cells.

BACKGROUND: HOXA10 is closely related to tumor progression in many human cancers. However, the role of HOXA10 in pancreatic cancer remains unclear. The aim of this study was to determine the involvement of HOXA10 in pancreatic cancer cell invasion and migration. METHODS: The effect of HOXA10 on the invasion and migration of pancreatic cancer cells was assessed by invasion and migration assays. The protein of transforming growth factor beta-2 (TGFß2) was neutralized by TGFß2 blocking antibody. The activation of p38 was inhibited by SB239063. RESULTS: HOXA10 could promote the invasion and migration of pancreatic cancer cells. Knockdown of HOXA10 decreased the expressions of TGFß2 and matrix metallopeptidase-3 (MMP-3) and suppressed the activation of p38. Conversely, overexpression of HOXA10 increased the levels of TGFß2 and MMP-3. Further experiments identified that TGFß2 contributed to the HOXA10-promoted invasion and migration and regulated MMP-3 expression and p38 activation. Additionally, inhibition of p38 suppressed cell invasion and MMP-3 expression in pancreatic cancer cells. CONCLUSIONS: HOXA10 promotes cell invasion and MMP-3 expression of pancreatic cancer cells via TGFß2-p38 MAPK pathway. Thus, HOXA10 could be a useful target for the treatment of pancreatic cancer.

A multi-center investigation of breast-conserving surgery based on data from the Chinese Society of Breast Surgery (CSBrS-005).

BACKGROUND: Although breast-conserving surgery is one of the standard treatments for breast cancer, few studies have assessed its recent implementation in China. We aimed to clarify the current real-world status of breast-conserving surgery in China. METHODS: This cross-sectional survey relied on data collected by the Chinese Society of Breast Surgery (CSBrS) to examine patients who underwent this surgery between January 2018 and December 2018. The survey was conducted using a uniform electronic questionnaire to collect information, including clinical and pathological data on these patients. RESULTS: Overall, 4459 breast-conserving surgeries were performed in 34 member units of CSBrS, accounting for 14.6% of all breast cancer surgeries performed in these units during the study period. In patients who underwent breast-conserving surgery with information on tumor size available, more than half (61.2%) of the tumors were smaller than 2 cm in diameter, and only 87 (3.2%) tumors were larger than 4 cm in diameter. Among patients who underwent breast-conserving surgeries, 457 (10.2%) patients received neoadjuvant therapy before the surgery. Among patients with a reported margin width, 34 (2.0%) patients had a margin of ≤2 mm, and 1530 (88.2%) of them had a margin of >5 mm. CONCLUSIONS: This study demonstrated the rates of breast-conserving surgery in member units of the CSBrS, and introduced the characteristics and surgical margins of patients who underwent this surgery. This information helps describe the real-world status of breast-conserving surgery in China. TRIAL REGISTRATION: chictr.org.cn, ChiCTR1900026841; http://www.chictr.org.cn/showproj.aspx?proj=42783.

Clinicopathologic and prognostic characteristics of triple-negative breast cancer.

BACKGROUND: Triple-negative breast cancer (estrogen receptor (ER)-, progesterone receptor (PR)-, and HER2-negative) is a rare subtype with a poor prognosis. However, the clinicopathologic and prognostic characteristics of triple-negative breast cancer remain undetermined. MATERIALS AND METHODS: Immunohistochemical staining was adopted to examine the expressions of ER, PR, p53, C-erbB-2 (HER2), vascular endothelial growth factor (VEGF), and epidermal growth factor receptor (EGFR) protein in 116 samples of paraffin-embedded breast cancer tissues. RESULTS: 22 triple-negative breast cancers were found among 116 informative cases (19%). The triple-negative phenotype significantly correlates with tumor size, histological grade, lymph node status, p53, and EGFR (p < 0.05), and not significantly with age, menopausal status, and VEGF protein. After a median follow-up period of 96 months (range: 32-123 months), 12 triple-negative breast cancer patients and 20 patients with non-triple-negative phenotype had distant relapse (p < 0.05). Survival analysis showed that triple-negative phenotype was inversely associated with overall survival (p < 0.05) but not significantly with disease-free survival (p = 0.2877). Multivariate Cox model analysis showed that tumor size, lymph node status, histological grade, and triple-negative phenotype provided independent significant predictive power. CONCLUSION: Triple-negative breast cancer phenotype has specific clinical and biological characteristics. Patients with triple-negative breast cancer have a poorer prognosis. So far, there is no conclusive effective treatment, which necessitates further studies.

Six cases of chylous leakage after axillary lymph node dissection.

BACKGROUND: Chylous leakage has been described after several surgical procedures, especially in the region of the neck and thorax. However, it has rarely been reported after axillary lymph node dissection. PATIENTS AND METHODS: We encountered 6 cases of chylous leakage after axillary lymph node dissection out of a total of 882 breast cancer patients between July 2005 and June 2007 in Shandong Provincial Hospital. These 6 cases were confirmed by axillary white fluid and chylomicron interpretation. The patients were treated conservatively, including a low fat diet, compression bandage, and suction drainage. RESULTS: All 6 cases were successfully treated without any complications such as infection, dystrophy, and lymphoceles. The chylous leakage disappeared within a median of 5 days (range: 3-7 days). Adjuvant chemotherapy and radiotherapy were not delayed. After a median follow-up period of 12 months (range: 6-20 months), no chronic complications were observed. CONCLUSION: Chylous leakage after axillary lymph node dissection is quite rare. It can be cured by conservative treatment. Lymphatic vessels should be identified carefully, and the main duct should be carefully ligated during surgical procedures, especially when level II and III lymph nodes are removed.

[Inhibitive effects of mifepristone on growth of breast cancer: experiment with animal model].

OBJECTIVE: To investigate the effects of mifepristone (MIF) on the growth of breast cancer. METHODS: Forty female athymic BALB/c-nude mice underwent subcutaneous injection of breast cancer cells of the line MCF-7, ER +/PR +. Ten days later when tumor nodules were formed, the mice were randomly divided into 4 equal groups to be administered with MIF of the concentrations of 25 mg, 50 mg, 100 mg, and 200 mg/kg x d respectively by gastric perfusion. The tumor size was observed every 3 day till 3 weeks later. Ten mice were used as normal control group, undergoing gastric perfusion of vegetable oil. Parts of the animals were killed 2 weeks later, and the remaining mice were all killed 3 weeks later. The tumors were taken out and underwent immunochemistry to measure the protein expression of CD34, estrogen receptor (ER), progesterone receptor (PR), vascular endothelial growth factor (VEGF), bcl-2, Ki67, p53, and CerbB-2. Microscopy was used to measure the microvessel density (MVD). RESULTS: The growth velocity of tumor of the mice of MIF groups were all slower than that of the control group (all P <0.01). The MVD levels of the MIF groups all decreased time- and dose-dependently. Microscopy showed that in the tumor tissues heteromorphism was significant, pathological caryomitosis was more remarkable, karyoplasmic ratio was greater, endochylema was deep blue, mesenchyma was sparse, and zone of neoplasm necrosis became lager in comparison with the control group. The expression levels of VEGF, bcl-2, Ki67, p53, and CerbB-2 of the MIF groups were all significantly lower, time and dose-dependently, than those of the control group (all P <0.05). CONCLUSION: MIF inhibits the growth of breast cancer by the mechanisms related to apoptosis promotion and inhibition of angiogenesis, so it can be used in breast cancer endocrine therapy.

Crosstalk analysis of pathways in breast cancer using a network model based on overlapping differentially expressed genes.

Multiple signal transduction pathways can affect each other considerably through crosstalk. However, the presence and extent of this phenomenon have not been rigorously studied. The aim of the present study was to identify strong and normal interactions between pathways in breast cancer and determine the main pathway. Five sets of breast cancer data were downloaded from the high-throughput Gene Expression Omnibus (GEO) and analyzed to identify differentially expressed (DE) genes using the Rank Product (RankProd) method. A list of pathways with differential expression was obtained by gene set enrichment analysis (GSEA) of the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The DE genes that overlapped between pathways were identified and a crosstalk network diagram based on the overlap of DE genes was constructed. A total of 1,464 DE genes and 26 pathways were identified. In addition, the number of DE genes that overlapped between specific pathways were determined, and the greatest degree of overlap was between the extracellular matrix (ECM)-receptor interaction and Focal adhesion pathways, which had 22 overlapping DE genes. Weighted pathway analysis of the crosstalk between pathways identified that Pathways in cancer was the main pathway in breast cancer.

Correlation between the expression of S100A4 and the efficacy of TAC neoadjuvant chemotherapy in breast cancer.

The aim of this study was to investigate the correlation between the expression of S100A4 and the efficacy of neoadjuvant chemotherapy in breast cancer. A total of 65 patients with invasive breast cancer were treated with neoadjuvant chemotherapy using the TAC regimen. The expression of S100A4 was detected by an immunohistochemical two-step method prior to treatment, after 2 cycles of chemotherapy and after 4 cycles of chemotherapy. Pathological evaluations of the chemotherapy were performed using the Miller and Payne (MP) grading system and their correlation with the changes of S100A4 expression during and after the treatment were explored. Between pre-neoadjuvant chemotherapy and 4 cycles post-chemotherapy, there was a significant difference in the expression of S100A4 (P<0.05); S100A4 expression was associated with neoadjuvant chemotherapy. However, between pre-neoadjuvant chemotherapy and 2 cycles post-chemotherapy, there was no significant difference in the expression of S100A4 (P>0.05). The intensity and changes of S100A4 expression were positively correlated with the efficacy of neoadjuvant chemotherapy (r=0.259, P<0.05). When patients with an MP grade of I or II following the second cycle of neoadjuvant chemotherapy were continually treated with the original chemotherapy for another 2 cycles, the desired effect was generally not achieved. S100A4 may be used as a predictor of the efficacy of neoadjuvant chemotherapy in breast cancer, guiding the formulation of individualized programs to improve the effectiveness of the treatment. For patients with an MP grade level of I or II after 2 cycles of neoadjuvant chemotherapy, the use of alternative chemotherapy regimens should be considered.

ABT-737 reverses the acquired radioresistance of breast cancer cells by targeting Bcl-2 and Bcl-xL.

BACKGROUND: Acquired radioresistance of cancer cells remains a fundamental barrier to attaining the maximal efficacy of radiotherapy for the treatment of breast cancer. Anti-apoptotic proteins, such as Bcl-2 and Bcl-xL, play an important role in the radioresistance of cancer cells. In the present study, we aimed to determine if ABT-737, a BH3-only mimic, could reverse the acquired radioresistance of the breast cancer cell line MDA-MB-231R by targeting Bcl-2 and Bcl-xL. METHODS: The radiosensitivity of MDA-MB-231 and MDA-MB-231R cells was compared using colony formation assays. Reverse-transcription PCR and western blot were performed to detect the expression of Bcl-2 and Bcl-xL in the cancer cell lines. Annexin V flow cytometric analysis and caspase-3 colorimetric assay were used to evaluate apoptosis of the cancer cells. Cell viability was measured using the Cell Counting Kit-8. The animals used in this study were 4 to 6-week-old athymic female BALB/c nu/nu mice. RESULTS: The MDA-MB-231R cells were more radioresistant than the MDA-MB-231 cells, and Bcl-2 and Bcl-xL were overexpressed in the MDA-MB-231R cells. While ABT-737 was able to restore the radiosensitivity of the MDA-MB-231R cells in vitro and in vivo experiment, it was not able to enhance the radiosensitivity of the MDA-MB-231 cells. In addition, ABT-737 increased radiation-induced apoptosis in the MDA-MB-231R cells. Bcl-2 and Bcl-xL were down regulated in the MDA-MB-231R cells following treatment with ABT-737. CONCLUSIONS: Targeting of the anti-apoptotic proteins Bcl-2 and Bcl-xL with ABT-737 may reverse the acquired radioresistance of MDA-MB-231R cells in vitro and in vivo. These findings suggest an attractive strategy for overcoming the acquired radioresistance of breast cancer cells.

Elevated thymidine kinase 1 in serum following neoadjuvant chemotherapy predicts poor outcome for patients with locally advanced breast cancer.

Patients with locally advanced breast cancer (LABC) commonly have an unfavorable prognosis. A molecular predictor for the identification of at-risk patients is urgently required. Thymidine kinase 1 in serum (S-TK1) is an enzyme involved in the synthesis of DNA precursors. In studies using immunohistochemistry, it was reported to be a more useful proliferation marker than Ki-67 in breast, lung and colorectal carcinoma. In the present study, we extended the research of prior breast carcinoma studies by postulating that in patients with LABC, overexpression of S-TK1 following neoadjuvant chemotherapy predicts cancer outcome. An experimental design consisting of 48 patients with LABC was prospectively constructed and analyzed. All patients received neoadjuvant chemotherapy and definitive surgical therapy. Study homogeneity was maintained by standardized treatment, surveillance and compliance protocols. The S-TK1 concentration was detected using the anti-TK1 chicken IgY antibody, using a dot-blot immuno-assay. After a median follow-up of 30 months, the results indicated a statistically significant trend (unadjusted). Patients with high S-TK1 overexpression had a significantly higher incidence of recurrence (P=0.006) and cancer death (P= 0.0128) than those with low S-TK1 overexpression. A multivariate analysis provided identical results. The hazards ratio for developing recurrence in patients with higher S-TK1 expression was 6-7 times higher than the hazards ratio in patients with lower expression. In conclusion, our results indicate that a high S-TK1 concentration in sera from LABC patients receiving neoadjuvant chemotherapy is predictive of cancer outcome.