Prognostic factors for breast cancer patients with T1-2 tumors and 1-3 positive lymph nodes and the role of postmastectomy radiotherapy in these patients.

PURPOSE: The purpose of this study was to identify independent prognostic factors for breast cancer patients with T1-2 tumors and 1-3 positive lymph nodes, and discuss the role of postmastectomy radiotherapy(PMRT) in these patients. METHODS: Between January 2005 and December 2015, the data on 840 eligible patients with breast cancer were retrospectively reviewed. Of these patients, 368 women received PMRT and 472 did not. The endpoints were locoregional recurrence (LRR) and distant metastasis (DM). RESULTS: With a median follow-up of 62.0 months, multivariate analysis identified the following independent risk factors for increased LRR: tumor size ≥ 4 cm (HR: 2.994, 95% CI: 1.190-7.535, P = 0.020), ER- and PR-negative tumor (HR: 2.540, 95% CI: 1.165-5.537, P = 0.019), preoperative high neutrophil-to-lymphocyte ratio (NLR) (HR: 4.716, 95% CI: 1.776-12.528, P = 0.002)and low neutrophil-to-monocyte ratio (NMR) (HR: 0.231, 95% CI: 0.084-0.633, P = 0.004). And independent risk factors for increased DM: ER- and PR-negative tumor (HR: 2.540, 95% CI: 1.880-5.625, P = 0.000), high NLR (HR: 2.693, 95% CI: 1.426-5.084, P = 0.002) and low NMR (HR: 0.460, 95% CI: 0.257-0.824, P = 0.009). The high-risk patients (≥ 2 risk factors) had worse LRRFS and DFS than low-risk patients (0-1 risk factor) (all, P < 0.05). In the subgroup analysis, both low- and high-risk patients received PMRT had better LRRFS and DFS than those who without PMRT (all, P < 0.05), and the high-risk patients received PMRT had similar 5-year rates of LRRFS and DFS than low-risk patients who without PMRT (94.5 vs. 94.3%, P = 0.402; 83.4 vs.87.4%, P = 0.877, respectively). CONCLUSIONS: Tumor size, ER/PR status, preoperative NLR and NMR were independent predictors of risk of recurrence. PMRT could improve locoregional control even in low-risk subgroup of breast cancer patients with T1-2 tumors and 1-3 positive lymph nodes significantly.

Modification of Ti implant surface for cell proliferation and cell alignment.

Surface properties of implants are the keys for ensuring their long-lasting anchorage to the tissue. This study aims to develop a novel implant surface microstructure with high biocompatibility and ability of guided tissue formation. By a photolithography method, gold (Au) grids (1 x 1 mm(2) square lattices, 10 mum in grid-line width) were deposited on titanium substrates. They were oxidized with H(2)O(2) solution to yield titania (anatase) layer, and the Au grid formed channels due to larger molar volume of anatase than Ti. L-Cysteine and type I collagen were then immobilized on them to yield the target substrates, CHT-Au-cys-col. Apatite deposited within 3 days when they were soaked in Kokubo's simulated body fluid, regardless of the protein coating, but not on the bottom of the Au channel. Osteoblast-like MC3T3-E1 cells were cultured on the CHT-Au-cys-col substrates, showing that (1) the cysteine-collagen coating promoted cell attachment and proliferation, and (2) the Au channels were filled with the cells which were aligned along the channel direction and were connected to the neighboring cells as well as attached to the channel wall with cytoplasmic extensions. The results thus ensured filopodial guidance for the substrates.

[Cleavage of telomerase RNA component by two DNAzymes and their effects on expression of two apoptosis-related genes in human mammary cancer cells].

OBJECTIVE: To investigate the cleavage of telomerase RNA component (hTR) by two DNAzymes and their effects on the expression of two apoptosis-related genes in a human mammary cancer cell line (MCF-7). METHODS: Two "10-23" DNAzymes (DzT and DzTi) targeted against hTR RNA and their analogues (DzT' and DzTi') were synthesized and used to cleave hTR RNA in vitro and in MCF-7 cells. Reverse transcriptional (RT) PCR amplification of a hTR DNA fragment and telomerase activity assay by PCR-enzyme-linked immunosorbent assay (ELISA) were performed to assess the cleavage efficiency. The biological effect of the DNAzymes was evaluated by measuring the expression of two apoptosis-related genes (bax-2 and bax). RESULTS: The unmodified DNAzyme, DzT, and its modified form, DzTi, which had an 3'-inverted thymidine added, could effectively cleave hTR mRNA in vitro. After transfection into MCF-7 cells, DzTi exhibited more powerful cleavage ability than DzT, and down-regulated the activity of telomerase (P<0.05). However, neither DzTi nor DzT displayed significant effect on the growth of the cells (P>0.05) and the expression of bax-2 and bax (P>0.05). DzT' and DzTi', derived from DzT and DzTi respectively with a base displacement in the conserved catalytic motif, did not exhibit notable cleavage effect on hTR RNA either in vitro or in the cells. CONCLUSIONS: The synthesized DNAzymes can effectively and specifically cleave hTR RNA and decrease the telomerase activity. As a newly found catalytic nucleic acid, DNAzymes will play an important role in gene therapy studies of tumors.

Peroxisome proliferator-activated receptor gamma ligands inhibit cell growth and induce apoptosis in human liver cancer BEL-7402 cells.

AIM: To investigate the characteristics of PPAR gamma ligands induced apoptosis in liver cancer cells. METHODS: The effects of ligands for each of the PPAR gamma ligands on DNA synthesis and cell viability were examined in BEL-7402 liver cancer cells. Apoptosis was characterized by Hochest33258 staining, DNA fragmentation, TUNEL and ELISA, and cell cycle kinetics by FACS. Modulation of apoptosis related caspases expression by PPAR gamma ligands was examined by Western blot. RESULTS: PPARgamma ligands, 15-deoxy-(12), (14)-prostaglandin J2 (15d-PGJ2) and troglitazone (TGZ), suppressed DNA synthesis of BEL-7402 cells. Both 15d-PGJ2 and TGZ induced BEL-7402 cell death in a dose dependent manner, which was associated with an increase in fragmented DNA and TUNEL-positive cells. At concentrations of 10 and 30 microM, 15d-PGJ(2) or troglitazone increased the proportion of cells with G(0)/G(1) phase DNA content and decreased those with S phase DNA content. There was no significant change in the proportion of cells with G(2)/M DNA content. The activities of Caspases-3, -6, -7 and -9 were increased by 15d-PGJ2 and TGZ treatment, while the activity of Caspase 8 had not significantly changed. CONCLUSION: The present results suggest the potential usefulness of PPAR gamma ligands for chemoprevention and treatment of liver cancers.

PPARgamma pathway activation results in apoptosis and COX-2 inhibition in HepG2 cells.

AIM: To investigate whether troglitazone (TGZ), the peroxisome proliferator-activated receptor (PPAR) gamma ligand, can induce apoptosis and inhibit cell proliferation in human liver cancer cell line HepG2 and to explore the molecular mechanisms. METHODS: (3-(4,5)-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), ((3)H) Thymidine incorporation, Hochest33258 staining, DNA ladder, enzyme-linked immunosorbent assay (ELISA), RT-PCR, Northern and Western blotting analyses were employed to investigate the effect of TGZ on HepG2 cells and related molecular mechanisms. RESULTS: TGZ was found to inhibit the growth of HepG2 cells and to induce apoptosis. During the process, the expression of COX-2 mRNA and protein and Bcl-2 protein was down-regulated, while that of Bax and Bak proteins was up-regulated, and the activity of caspase-3 was elevated. Furthermore, the level of PGE(2) was decreased transiently after 12 h of treatment with 30 microM troglitazone. CONCLUSION: TGZ inhibits cell proliferation and induces apoptosis in HepG2 cells, which may be associated with the activation of caspase-3-like proteases, down-regulation of the expression of COX-2 mRNA and protein, Bcl-2 protein, the elevation of PGE2 levels, and up-regulation of the expressions of Bax and Bak proteins.