SQSTM1/p62 regulate breast cancer progression and metastasis by inducing cell cycle arrest and regulating immune cell infiltration.

The autophagy adaptor protein SQSTM1/p62 is overexpressed in breast cancer and has been identified as a metastasis-related protein. However, the mechanism by which SQSTM1/p62 contributes to breast cancer progression and tumor microenvironment remains unclear. This study revealed that silencing SQSTM1/p62 expression suppressed breast cancer progression via regulating cell proliferation and reshaping the tumor microenvironment (TME). Here, we found that SQSTM1/p62 was overexpressed in multiple human cancer tissue types and that was correlated with poor patient overall survival (OS) and disease-free survival (DFS). Moreover, we found that short-hairpin RNA (shRNA)-mediated knockdown of p62 expression significantly inhibited cell proliferation, migration, and invasion, and promoted cell death in vitro, as well as suppressed breast cancer growth and lung metastasis in vivo. In addition, flow cytometry analysis of splenocytes and tumor infiltrating lymphocytes (TILs) indicated that the numbers of CD8α+ interferon (IFN)-γ+ cells (CTLs) and CD4+IFN-γ+ (Th1) cells were increased while those of CD4+IL-4+ (Th2) cells, tumor-associated macrophages (TAMs) and myeloid-derived suppressor cells (MDSCs) were decreased. RT-PCR analyses showed that the gene expression of Th1/Th2 cytokines changed in the tumor microenvironment. Silencing SQSTM1/p62 suppressed tumor cell lung metastasis. Together, our results provide strong evidence that silencing tumor cell SQSTM1/p62 inhibited tumor growth and metastasis through cell cycle arrest and TME regulation. This finding provides a novel molecular therapeutic strategy for breast cancer progression and metastasis treatment.

Pulmonary Staphylococcus aureus infection regulates breast cancer cell metastasis via neutrophil extracellular traps (NETs) formation.

The formation of neutrophil extracellular traps (NETs) was recently identified as one of the most important processes for the maintenance of host tissue homeostasis in bacterial infection. Meanwhile, pneumonia infection has a poor effect on cancer patients receiving immunotherapy. Whether pneumonia-mediated NETs increase lung metastasis remains unclear. In this study, we identified a critical role for multidrug-resistant Staphylococcus aureus infection-induced NETs in the regulation of cancer cell metastasis. Notably, S. aureus triggered autophagy-dependent NETs formation in vitro and in vivo and increased cancer cell metastasis. Targeting autophagy effectively regulated NETs formation, which contributed to the control of cancer metastasis in vivo. Moreover, the degradation of NETs by DNase I significantly suppresses metastasis in lung. Our work offers novel insight into the mechanisms of metastasis induced by bacterial pneumonia and provides a potential therapeutic strategy for pneumonia-related metastasis.

P. aeruginosa Mediated Necroptosis in Mouse Tumor Cells Induces Long-Lasting Systemic Antitumor Immunity.

Necroptosis is a form of programmed cell death (PCD) characterized by RIP3 mediated MLKL activation and increased membrane permeability via MLKL oligomerization. Tumor cell immunogenic cell death (ICD) has been considered to be essential for the anti-tumor response, which is associated with DC recruitment, activation, and maturation. In this study, we found that P. aeruginosa showed its potential to suppress tumor growth and enable long-lasting anti-tumor immunity in vivo. What's more, phosphorylation- RIP3 and MLKL activation induced by P. aeruginosa infection resulted in tumor cell necrotic cell death and HMGB1 production, indicating that P. aeruginosa can cause immunogenic cell death. The necrotic cell death can further drive a robust anti-tumor response via promoting tumor cell death, inhibiting tumor cell proliferation, and modulating systemic immune responses and local immune microenvironment in tumor. Moreover, dying tumor cells killed by P. aeruginosa can catalyze DC maturation, which enhanced the antigen-presenting ability of DC cells. These findings demonstrate that P. aeruginosa can induce immunogenic cell death and trigger a robust long-lasting anti-tumor response along with reshaping tumor microenvironment.

[Comparison of two treatment method for maxillary incisors intrusion].

PURPOSE: To compare the clinical effects of miniscrews and conventional utility arch in the maxillary incisors intrusion. METHODS: Twenty adult patients with III degree deep bite were chosen and divided into two groups randomly. The cephalometric films, intraoral periapical radiographies with metal guide bars, and periodontium parameters were taken before and after intrusion. The major measurement items included alteration of the maxillary first molar in vertical dimension, torque and length of maxillary central incisors, overbite, clinical crown of maxillary central incisors, probing depth (PD), gingival index (GI), plaque index (PI) and etc. PASW statistics 18.0 was used for statistical analysis. RESULTS: In the miniscrew group, there was no change of U1-SN, U6-PP distance, U6-SN(P<0.05), but significant reductions in overbite and U1-Stm distance(P<0.05). The probing depth (PD) increased(P<0.05), and the width of keratinized gingiva(WKG) decreased significantly(P<0.05). In the conventional utility arch group, significant differences were found in U1-SN, U6-SN, OP-SN, PLI2(P<0.05). There was no difference in periodontal supporting tissue changes between the two groups(P>0.05). CONCLUSIONS: Compared with the conventional utility arch, minisrew has advantages on intruding upper incisors, and improving lip-tooth relationships. Less treatment period is needed in the minisrew group, and the impact on periodontal supporting tissue is similar.

Effects of traditional Chinese medicine on bone remodeling during orthodontic tooth movement.

ETHNOPHARMACOLOGICAL RELEVANCE: Dipsacus asper Wall (Dipsacaceae), Salvia miltiorrhiza (Salvia) and Drynaria fortunei (Drynaria) have been traditionally used in Chinese medicine as the main ingredient of many formulations for the treatment of cardiovascular and inflammatory diseases. AIM OF THE STUDY: This study was aimed to evaluate the effect of the traditional Chinese medicine (TCM) Dipsacaceae, Salvia and Drynaria on bone remodeling during orthodontic tooth movement (OTM). SETTING: This study was conducted in School of Stomatology, Shandong University between 2009 and 2010 [Jinan, Shandong, China]. MATERIALS AND METHODS: Ninety-six eight-week-old female SPF Wistar rats 180-200 g were selected and randomly divided into four groups of 24: Dipsacaceae group, Salvia group, Drynaria group and control group. Animal models for orthodontic tooth movement were then established which consisted of a closed coil spring ligated to the upper first molar and incisors, exerting a force of 40 g during the experimental period. Rats in the TCM groups were given Dipsacaceae, Salvia and Drynaria decoction respectively by intragastric administration 6 g/kg/day and the control group were given normal saline 3 ml. The rats were sacrificed in batch on the 7th, 14th, 21st and 28th days after orthodontic treatment. Slices from periodontium of the upper first molar were observed under optical microscope. Neovascularization, new bone formation and osteoclast number were observed. RESULTS: The upper first molars were drawn mesial by the force. Telangiectasia and new bone formation in periodontal tissue were significantly in the TCM groups compared with the control group. Application of orthodontic forces in the experimental teeth showed a significant increase (P<0.05) of osteoclast number in the TCM group when compared with the control group. In addition, the number of osteoclast had no significant differences among the TCM groups (P>0.05). Osteoclast number in the TCM group and the control group were 10.12±0.058, 10.13±0.022, 10.09±0.047 and 9.55±0.045, respectively. CONCLUSIONS: These findings suggest that the TCM decoction are beneficial to the alveolar bone remodeling by promoting osteoclast differentiation during OTM.

[Detection of chromosomal DNA imbalance in medulloblastoma by comparative genomic hybridization].

OBJECTIVE: To investigate the relationship between chromosomal genomic DNA imbalance in medulloblastoma (MB), and the age and gender. METHODS: The gains and losses of chromosomal genomic DNA in 16 MBs were analyzed using comparative genomic hybridization. RESULTS: The gains and(or) losses were found in 15 of the 16 cases. There was not significant difference (P > 0.05) between the total gains (10/16) and losses (11/16). Both of their differences had also no significance between different age and gender groups (P > 0.05). In 15 cases with gains and(or) losses, single-, two-, three- and multi-chromosome genomic DNA imbalances were 3/15, 4/15, 1/15 and 7/15 respectively. Eleven gain zones (+5q, +6q, +7q, +11q, +15q, +17p, +17q, +19q, +20q, +21q, +Xp) and twenty-five loss zones (-1p, -1q, -2p, -2q, -3q, -4p, -6p, -6q, -8p, -8q, -10p, -10q, -11p, -14q, -16p, -16q, -17p, -18p, -18q, -19p, -19q, -20p, -20q, -Xp, -Xq) were detected in those tumors. +7q (6/16), +17q (6/16), -14q (5/16) and -10q (3/16) were the most frequent, but -14q only occurred in the cases of > 10-year-old. CONCLUSIONS: Most MBs have chromosomal genomic DNA imbalances. The frequent imbalance zones are mainly at the long arms of some chromosomes. +7q, +17q, -14q and -10q correlate closely to development of the tumors. -14q is important factor to result in MBs of > 10-year-old group. MB has possibly different molecular genetics subtype.

[The establishment of three dimensional finite element model of a maxillary impacted canine and its support tissues].

PURPOSE: To establish a three dimensional finite element model of a maxillary impacted canine and its support tissues by means of finite element method (FEM) and spiral CT which can offer digital model for orthodontic traction. METHODS: A patient with a completely bony impacted maxillary canine was chosen. The CAD model of the canine and its surrounding tissues was established using spiral CT equipment, Mimics and Unigraphic. The CAD model was changed to FEM models by MSC. Mentat which could cut nets in CAD models. RESULTS: With the use of spiral CT,Mimics,UG and MSC, FEM model of the maxillary impacted canine and its support tissues and the whole maxilla was constructed and the model was of high geometrical similarity. CONCLUSION: The FEM model of maxillary impacted canine established by spiral CT,Mimics,UG and MSC has high accuracy, integrated constitution and optimized mesh which can be taken into account in the course of the treatment planning prior to starting therapy.

[Detection of chromosomal imbalance in ependymoma by comparative genomic hybridization].

OBJECTIVE: To investigate genomic DNA imbalances in ependymomas (EDMs) and their correlations with the tumor histological types, grades, locations, patients' gender and age. METHODS: Chromosomal gains and losses in 16 cases of EDM were analyzed using comparative genomic hybridization. RESULTS: Chromosomal regional gain and loss were found in 15 and 13 of 16 EDM cases respectively including totally 24 regional gains and 19 regional losses in all the tumors studied. Both regional gains and losses were mostly seen in myxopapillary EDMs (MPE, WHO grade I), more commonly seen in cellular EDMs (CE, WHO grade II) and tanycytic EDMs (TE, WHO grade II) than in anaplastic EDMs (AE, WHO grade III). Some of the regional gains and losses appeared only in one subtype of MPE, CE, TE and AE cases resulting in development of specific imbalance profiles of certain subtype in these cases. MPE, CE and TE often had +7. Chromosomal +5 occurred only in MPE and CE, and -22q was only seen in CE and TE. AE frequently had +1q, but none had +5, +7, -4q, -19q and -22q. The frequencies of any regional gain or loss were not affected by patients' genders (P > 0.05). Chromosomal +1q and +7p happened predominantly in intracranial EDMs with an averagely onset age of

[Expression of MMPs in gingival crevicular fluid of the rats in different age groups with orthodontic appliances].

PURPOSE: To detect the changes of MMP-1 and MMP-9 in gingival crevicular fluid (GCF) of the rats in different age groups with orthodontic appliances. METHODS: 40 female Wistar rats were selected as the animal model, in which there were 20 6-week-old rats and 20 12-week-old rats. The orthodontic appliances were fixed onto the maxilla of the rats after general anesthesia. The maxillary first molar in the experimental side was drawn medially by applying an excess force of 50 g, with both of the maxillary central incisors as anchorage. In the control side, the maxillary first molar was winded around the dental cervix with ligature wire. The GCF was collected every week, and MMP-1 and MMP-9 were assayed in quantitation by SDS-PAGE, Western blot and Gel Doc quantitative analysis system,respectively. The data were analyzed for ANOVA using SPSS11.0 software package. RESULTS: No statistical difference of the original concentration of MMP-1 and MMP-9 in GCF was found between adolescent group and adult group (P>0.05), but the adult group's contents were a little more than adolescent group's, and is the same as the 1st week's content. The concentration of MMPs in adult group changed more greatly than that in adolescent group. At the beginning of the experiment, the MMPs' increasing speed of adult group was higher than that of adolescent group. But at the end, the rate of descent of adult group was lower than that of adolescent group. CONCLUSIONS: The expression of MMPs in gingival crevicular fluid of adult rats is higher than that of adolescent rats. It may be more serious of the orthodontic root resorption in adult rats than in adolescent rats. Supported by Shandong Provincial Natural Science Foundation (Grant No.Y2007C023) and Jinan Science and Technology Development Plan (Grant No.200705089-9).