A novel polypeptide CAPG-171aa encoded by circCAPG plays a critical role in triple-negative breast cancer.

BACKGROUND: The treatment of Triple-negative breast cancer (TNBC) has always been challenging due to its heterogeneity and the absence of well-defined molecular targets. The present study aims to elucidate the role of protein-coding circRNAs in the etiology and carcinogenesis of TNBC. METHODS: CircRNA expression data in TNBC (GEO: GSE113230, GSE101123) were reanalyzed and then circCAPG was selected for further study. To identify the polypeptide-coding function of circCAPG, a series of experiments, such as Mass spectrometry and dual-luciferase reporter assays were conducted. Cell proliferation, apoptosis and metastasis parameters were determined to investigate the cancerous functions CAPG-171aa plays in both TNBC organoids and nude mice. Mechanistically, the relation between CAPG-171aa and STK38 in TNBC was verified by immunoprecipitation analyses and mass spectrometry. The interactions between SLU7 and its binding site on circCAPG were validated by RIP-qPCR experiments. RESULTS: In both TNBC clinical samples and cell lines, the expression level of circCAPG was identified to be higher compared with normal ones and positively correlated with the overall survival (n = 132) in a 10-year follow-up study, in which the area under the curve of receiver operating characteristic was 0.8723 with 100% specificity and 80% sensitivity. In addition, we found that circCAPG knockdown (KD) significantly inhibited the growth of TNBC organoids. Intriguingly, circCAPG can be translated into a polypeptide named CAPG-171aa which promotes tumor growh by disrupting the binding of serine/threonine kinase 38 (STK38) to SMAD-specific E3 ubiquitin protein ligase 1 (SMURF1) and thereby preventing MEKK2 ubiquitination and proteasomal degradation. Furthermore, we found that SLU7 Homolog- Splicing Factor (SLU7) can regulate the bio-generation of circCAPG through binding to the flanking Alu sequences of circRNA transcripts. CONCLUSIONS: circCAPG significantly enhances the proliferation and metastasis of TNBC cells by encoding a novel polypeptide CAPG-171aa and afterwards activates MEKK2-MEK1/2-ERK1/2 pathway. Additionally, the formation of circCAPG is found to be mediated by SLU7. The present study provides innovative insight into the role of protein-coding circRNAs CAPG-171aa in TNBC, and its capacity to serve as a promising prognostic biomarker and potential therapeutic target in TNBC.

circMRPS35 promotes malignant progression and cisplatin resistance in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the major causes of cancer-related death worldwide. Circular RNAs (circRNAs), a novel class of non-coding RNA, have been reported to be involved in the etiology of various malignancies. However, the underlying cellular mechanisms of circRNAs implicated in the pathogenesis of HCC remain unknown. In this study, we identified a functional RNA, hsa_circ_0000384 (circMRPS35), from public tumor databases using a set of computational analyses, and we further identified that circMRPS35 was highly expressed in 35 pairs of HCC from patients. Moreover, knockdown of the expression of circMRPS35 in Huh-7 and HCC-LM3 cells suppressed their proliferation, migration, invasion, clone formation, and cell cycle in vitro, and it suppressed tumor growth in vivo as well. Mechanically, circMRPS35 sponged microRNA-148a-3p (miR-148a), regulating the expression of Syntaxin 3 (STX3), which modulated the ubiquitination and degradation of phosphatase and tensin homolog (PTEN). Unexpectedly, we detected a peptide encoded by circMRPS35 (circMRPS35-168aa), which was significantly induced by chemotherapeutic drugs and promoted cisplatin resistance in HCC. These results demonstrated that circMRPS35 might be a novel mediator in HCC progress, and they raise the potential of a new biomarker for HCC diagnosis and prognosis, as well as a novel therapeutic target for HCC patients.

Improvement of RTT Fairness Problem in BBR Congestion Control Algorithm by Gamma Correction.

Google proposed the bottleneck bandwidth and round-trip propagation time (BBR), which is a new congestion control algorithm. BBR creates a network path model by measuring the available bottleneck bandwidth and the minimum round-trip time (RTT) to maximize delivery rate and minimize latency. However, some studies have shown that there are serious RTT fairness problems in the BBR algorithm. The flow with longer RTT will consume more bandwidth and the flows with shorter RTT will be severely squeezed or even starved to death. Moreover, these studies pointed out that even small RTT differences will lead to the throughput of BBR flows being unfair. In order to solve the problem of RTT fairness, an improved algorithm BBR-gamma correction (BBR-GC) is proposed. BBR-GC algorithm takes RTT as feedback information, and then uses the gamma correction function to fit the adaptive pacing gain. This approach can make different RTT flows compete for bandwidth more fairly, thus alleviating the RTT fairness issue. The simulation results of Network Simulator 3 (NS3) show that that BBR-GC algorithm cannot only ensure the channel utilization, but also alleviate the RTT fairness problem of BBR flow in different periods. Through the BBR-GC algorithm, RTT fairness is improved by 50% and the retransmission rate is reduced by more than 26%, compared with that of the original BBR in different buffer sizes.

PFKFB4 Promotes Breast Cancer Metastasis via Induction of Hyaluronan Production in a p38-Dependent Manner.

BACKGROUND/AIMS: The bi-functional enzyme 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase-4 (PFKFB4) is highly expressed in many types of cancer and its requirement for tumor survival has been demonstrated in glioma, lung, and prostate cancers. However, whether PFKFB4 plays a role in the tumor metastasis remains uncertain. This study explores the role of PFKFB4 in tumor metastasis and its underlying mechanisms in breast cancer cells. METHODS: The expression of PFKFB4 was first analyzed using the Cancer Genome Atlas (TCGA) dataset, and confirmed by immunohistochemical staining of tissue microarray and breast cancer tissues from patient samples. Gain- and loss-of- function approaches were used to investigate the effects of PFKFB4 on breast cancer cell migration in vitro. Orthotopic xenograft model and experimental metastasis model were used to assess the effects of PFKFB4 on breast cancer cell metastasis in vivo. ELISA and immunofluorescence staining were used to examine HA production. Quantitative RT-PCR and western blotting were used to explore the mRNA and protein levels of HAS2, respectively. RESULTS: We found that PFKFB4 enhances the migration/invasiveness of breast cancer cells in vitro as well as in vivo. Notably, the effects of PFKFB4 on migration are mediated by induction of HAS2 expression and HA production. Moreover, PFKFB4-induced HAS2 up-regulation depends upon the activation of p38 signaling. CONCLUSION: PFKFB4 promotes the metastasis of breast cancer cells via induction of HAS2 expression and HA production in a p38-dependent manner. Therefore, the PFKFB4/p38/HAS2 signaling pathway may serve as a potential therapeutic target for metastatic breast cancer.

Comparison of surgical complication between immediate implant and autologous breast reconstruction after mastectomy: A multicenter study of 426 cases.

BACKGROUND AND OBJECTIVES: There is a lack of multicenter immediate breast reconstruction data comparing the surgical complication of implant and autologous breast reconstruction, especially in China. In this study, we used the data from eight centers to study the complications and their risk factors in this population. METHODS: Sociodemographic and clinicopathological data were obtained and compared for patients who received immediate implant and autologous breast reconstruction after breast cancer surgery in the eight hospitals between 2012 and 2016. Logistic regression analysis was used to identify risk factors associated with the complication of breast reconstruction. RESULTS: Immediate autologous reconstruction (IAR) was associated with significantly higher rates of overall complications (P = 0.036), fat liquefaction (P < 0.001), and reconstructive failure (P = 0.019), but lower rates of wound complications (P = 0.01) compared with the immediate implant reconstruction (IIR) at the median follow-up time of 13.6 months. With the logistic regression analysis, older patient (odds ratio [OR], 2.22; 95% confidence interval [CI], 1.15-4.28; P = 0.017), and obesity (OR, 2.17; 95% CI, 1.08-4.37; P = 0.030) were significant predictors of increased complications. CONCLUSION: Our multicenter results demonstrated that the rates of overall complications and reconstruction failure were higher after IAR than IIR. These findings can be used to better help surgeons and their patients with objective and reliable information to assist in selecting the modality of reconstruction.

Rosai-Dorfman disease with features of IgG4-related disease in the breast: Cases report and literature review.

BACKGROUND: A proportion of cases of Rosai-Dorfman disease exhibit some histological features consistent with IgG4-related disease (IgG4RD). Several investigators have discussed whether Rosai-Dorfman disease belongs to the spectrum of IgG4RD or is concurrent with it by coincidence. OBJECTIVE: To elucidate the relationship between the two diseases, we report key features, including IgG4 and amyloid levels, of four cases of Rosai-Dorfman disease in the breast. METHODS: The histological features of the four cases were analyzed and the numbers of IgG4+ plasma cells and IgG4/IgG ratios were evaluated. Serum IgG4 concentrations were also measured in two recent cases. A literature review was also performed. RESULTS: Two cases (case 1 and 2) showed features of IgG4RD, including lymphoid follicle formation with regressive changes, obliterative phlebitis, increased number of IgG4+ plasma cells, and increased IgG4/IgG ratio; one of the two had an elevated serum IgG4 level. Amyloidosis was detected in these cases, with amyloid in the stroma and the vessel walls of the lesion. The other two cases (case 3 and 4) only had mild increases in the numbers of IgG4+ plasma cells, while amyloid was deposited in the stroma only. CONCLUSIONS: A subset of Rosai-Dorfman disease may overlap with IgG4RD in the breast. When Rosai-Dorfman disease has features of IgG4RD, amyloidosis could be induced in the lesion.

Treatment patterns for adjuvant docetaxel-based chemotherapy in early-stage breast cancer in China: A pooled retrospective analysis of four observational studies.

OBJECTIVE: Adjuvant docetaxel-based chemotherapy is frequently used for operable early breast cancer (EBC). This study investigated patterns of use of docetaxel (T) in real-life clinical practice in China. METHODS: This was a retrospective pooled analysis of the Asia-Pacific Breast Initiatives (APBI) I (2006-2008) and II (2009-2011) registries, and two Chinese observational studies; BC STATE (2011-2014) and BC Local Registry (2007-2010). Female Chinese adults (≥18 years) with operable breast cancer treated with docetaxel-based adjuvant chemotherapy were included in the analysis. Patients with metastatic disease were excluded. The primary endpoint was assessment of treatment patterns and patient profiles. A logistic regression analysis was conducted to identify factors associated with choice of adjuvant chemotherapy regimen. RESULTS: Data from 3,020 patients were included. The most frequently used adjuvant regimen was docetaxel/anthracycline combination [n=1,421 (47.1%); of whom 52.0% received T/epirubicin (E)/cyclophosphamide (C)], followed by docetaxel/other [n=705 (23.3%); of whom 72.8% received TC], docetaxel/anthracycline sequential [n=447 (14.8%); of whom 40.9% and 39.6% received 5-Fu/EC-T and EC-T, respectively], and " other" [n=447 (14.8%); of whom 91.5% received T]. A significant association was found between adjuvant therapy with docetaxel/anthracycline combination and patient weight, menopausal status and estrogen receptor status. CONCLUSIONS: Real-world data revealed that docetaxel/anthracycline combination is the most commonly used category of docetaxel-based adjuvant therapy for patients with operable breast cancer in China; of which TEC is the most frequently used regimen.

Melatonin alleviates acute lung injury through inhibiting the NLRP3 inflammasome.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are clinically severe respiratory disorders, and there are currently no Food and Drug Administration-approved drug therapies. Melatonin is a well-known anti-inflammatory molecule, which has proven to be effective in ALI induced by many conditions. Emerging studies suggest that the NLRP3 inflammasome plays a critical role during ALI. How melatonin directly blocks activation of the NLRP3 inflammasome in ALI remains unclear. In this study, using an LPS-induced ALI mouse model, we found intratracheal (i.t.) administration of melatonin markedly reduced the pulmonary injury and decreased the infiltration of macrophages and neutrophils into lung. During ALI, the NLRP3 inflammasome is significantly activated with a large amount of IL-1ß and the activated caspase-1 occurring in the lung. Melatonin inhibits the activation of the NLRP3 inflammasome by both suppressing the release of extracellular histones and directly blocking histone-induced NLRP3 inflammasome activation. Notably, i.t. route of melatonin administration opens a more efficient therapeutic approach for treating ALI.

Oestrogen action and male fertility: experimental and clinical findings.

A proper balance between androgen and oestrogen is fundamental for normal male reproductive development and function in both animals and humans. This balance is governed by the cytochrome P450 aromatase, which is expressed also under spatio-temporal control. Oestrogen receptors ERα and/or ERß, together with the membrane-associated G-protein-coupled functional ER (GPER), mediate the effects of oestrogen in the testis. Oestrogen action in male reproduction is more complex than previously predicted. The androgen/oestrogen balance and its regulation in the masculinisation programming window (MPW) during foetal life is the most critical period for the development of the male reproductive system. If this balance is impaired during the MPW, the male reproductive system may be negatively affected. Recent data from genetically modified mice and human infertile patients have shown that oestrogens may promote the engulfment of live Leydig cells by macrophages leading to male infertility. We also discuss recent data on environmental oestrogen exposure in men and rodents, where a rodent-human distinction is crucial and analyse some aspects of male fertility potentially related to impaired oestrogen/androgen balance.

[Association between the expression of IGF1R and estrogen receptor and efficacy of neoadjuvant chemotherapy in beast cancer patients].

OBJECTIVE: To detect the expression of IGF1R and estrogen receptor, and to explore the relationship between their expression and the pathological complete response (pCR) rate of neoadjuvant chemotherapy (docetaxel plus epirubicin) in breast cancer patients. METHODS: We selected 139 women with breast cancer who underwent neoadjuvant chemotherapy (docetaxel plus epirubicin), and detected the expression of IGF1R and estrogen receptor in the samples taken before chemotherapy by Immunohistochemistry. The association between their expression and pCR rate of neoadjuvant chemotherapy was analyzed. RESULTS: Among the 139 cases, IGF1R was highly expressed in 45.3% (63/139) cases, and ER was positively expressed in 62.6% (87/139) cases. IGF1R was highly expressed in 54.0% (47/87) of the ER+ cases, significantly higher than that of ER- cases (30.8%, P<0.01). The overall pCR rate of all the 139 patients who received docetaxel plus epirubicin as neoadjuvant chemotherapy was 10.1% (14/139). The pCR rate was 19.2% (10/52) of the ER- patients and 4.6% (4/87) of the ER+ patients (P<0.05). The pCR rate was 10.5% (8/76) in the patients with low IGF1R expression and 9.5% (6/63) in the patients with high IGF1R expression (P>0.05). The patients with negative expression of ER and high expression of IGF1R showed the highest pCR rate (31.2%, P<0.01). CONCLUSIONS: Breast cancer patients with negative expression of ER and high expression of IGF1R are more sensitive to neoadjuvant chemotherapy of docetaxel plus epirubicin, and their pCR rate is significantly higher than that of other patients.

SOX2 regulates apoptosis through MAP4K4-survivin signaling pathway in human lung cancer cells.

Previous studies have implicated cancer stem cells in tumor recurrence and revealed that the stem cell gene SOX2 plays an important role in the tumor cell resistance to apoptosis. Nonetheless, the mechanism by which SOX2 regulates apoptosis signals remained undefined. Here, we demonstrated the surprising finding that silencing of the SOX2 gene effectively induces apoptosis via the activation of death receptor and mitochondrial signaling pathways in human non-small cell lung cancer cells. Unexpectedly, reverse transcription-PCR analysis suggested that downregulation of SOX2 leads to activation of MAP4K4, previously implicated in cell survival. Evaluation of the apoptotic pathways revealed an increased expression of key inducers of apoptosis, including tumor necrosis factor-α and p53, with concurrent attenuation of Survivin. Although p53 appeared dispensable for this pathway, the loss of Survivin in SOX2-deficient cells appeared critical for the observed MAP4K4 induced cell death. Rescue experiments revealed that SOX2-silencing-mediated killing was blocked by ectopic expression of Survivin, or by reduction of MAP4K4 expression. Clinically, expressions of Survivin and SOX2 were highly correlated with each other. The results reveal a key target of SOX2 expression and highlight the unexpected context-dependent role for MAP4K4, a pluripotent activator of several mitogen-activated protein kinase pathways, in regulating tumor cell survival.

Manufacture and evaluation of nano-silver-poly-DL-lactide-co-caprolactone-small intestinal submucosa biological mesh in abdominal wall reconstruction.

Background: Inguinal hernia repair is a routine surgical procedure and many methods are being applied to improve the operation. In this study, an abdominal wall defect model was established in New Zealand rabbits. The safety and efficacy of the test product was evaluated by observing the physiological state and clinical manifestations and conducting anatomical observations in the rabbits after use of the nano-silver-poly-DL-lactide-co-caprolactone-small intestinal submucosa (NS-PLCL-SIS) mesh. Methods: A total of 18 New Zealand rabbits were randomly divided into a test group and a blank group. Routine blood and serum biochemical tests, and anatomical observations were conducted on postoperative day 30 (D30), day 60 (D60), and day 90 (D90). During the study period, all animals underwent clinical observation, and the obtained data were counted. Results: The results showed that the NS-PLCL-SIS mesh was degraded within 90 days, and there was no abnormal reaction and no animal death during the test. There was no significant difference in the changes of animal body weight at each time point. There was no infectious inflammatory reaction in the wound at the study site, and the ocular wound healed well 7 days after the operation. Conclusions: Under the conditions of this experiment, the NS-PLCL-SIS mesh had good performance in the repair of abdominal wall defect in New Zealand rabbits and is clinically safe for veterinarians.

Tumor-derived mesenchymal progenitor cell-related genes in the regulation of breast cancer proliferation.

Background: Breast cancer (BC) is one of the most common malignancies worldwide, and its development is affected in various ways by the tumor microenvironment (TME). Tumor-derived mesenchymal progenitor cells (MPCs), as the most important components of the TME, participate in the proliferation and metastasis of BC in several ways. In this study, we aimed to characterize the genes associated with tumor-derived MPCs and determine their effects on BC cells. Methods: Tumor-derived MPCs and normal breast tissue-derived mesenchymal stem cells (MSCs) were isolated from tissues specimens of patients with BC. We conducted culture and passage, phenotype identification, proliferation and migration detection, inflammatory factor release detection, and other experiments on isolated MPCs from tumors and MSCs from normal breast tissues. Three paired tumor-derived MPCs and normal breast tissue-derived MSCs were then subjected to transcriptome analysis to determine the expression profiles of the relevant genes, and quantitative real-time polymerase chain reaction (qRT-PCR) was used to further confirm gene expression. Subsequently, the overexpression plasmids were transfected into tumor-derived MPCs, and the expression of various inflammatory factors of tumor-derived MPCs and their proliferation were characterized with a cell viability test reagent (Cell Counting Kit 8). Subsequently, the transfected tumor-derived MPCs were cocultured with BC cells using a conditioned medium coculture method to clarify the role of tumor-derived MSCs in BC. Results: Tumor-derived MPCs expressed stem cell characteristics including CD105, CD90, and CD73 and exhibited adipogenic and osteogenic differentiation in vitro. The proliferation of tumor-derived MPCs was significantly lower than that of normal breast tissue-derived MSCs, and the invasive metastatic ability was comparable; however, MPCs were found to release inflammatory factors such as interleukin 6 (IL-6) and transforming growth factor ß (TGF-ß). Transcriptome analysis showed that stomatin (STOM), collagen and calcium binding EGF domains 1 (CCBE1), and laminin subunit alpha 5 (LAMA5) were significantly upregulated in tumor-derived MPCs. Among them, STOM was highly expressed in tumor-derived MPCs, which mediated the slow proliferation of MPCs and promoted the proliferation of BC cells. Conclusions: STOM, CCBE1, and LAMA5 were highly expressed in tumor-derived MPCs, with STOM being found to retard the proliferation of MPCs but promote the proliferation of BC cells. There findings present new possibilities in targeted microenvironmental therapy for BC.

Optimization of apparent diffusion coefficient measured by diffusion-weighted MRI for diagnosis of breast lesions presenting as mass and non-mass-like enhancement.

The purpose of this study was to investigate the diagnostic value of the apparent diffusion coefficient (ADC), measured by diffusion-weighted magnetic resonance imaging (MRI), for the diagnosis of breast lesions presenting as mass and non-mass-like enhancement (NMLE). The breast MRI studies of 174 patients were reviewed retrospectively. A total of 188 histologically confirmed lesions were analyzed and classified into 127 mass enhancement (86 malignant and 41 benign) and 61 NMLE (42 malignant and 19 benign). The ADC values were measured using a spin-echo echo-planner-imaging (SE-EPI) sequence with b=1,000 s/mm(2). Diagnostic performance was evaluated using receiver operating characteristic (ROC) analysis. The mean ADC was 0.99 ± 0.22 × 10(-3)mm(2)/s for invasive cancer, 1.23 ± 0.33 × 10(-3)mm(2)/s for ductal carcinoma in situ (DCIS), and 1.52 ± 0.35 × 10(-3)mm(2)/s for benign adenosis. The mean ADC of all NMLE lesions was 1.44 ± 0.41 × 10(-3)mm(2)/s, which is higher than the mean ADC of all mass lesions, 1.12 ± 0.33 × 10(-3)mm(2)/s. In the ROC analysis, the optimal cutoff ADC value for differentiating benign from malignant lesions was 1.05 × 10(-3)mm(2)/s for mass lesions and 1.35 × 10(-3)mm(2)/s for NMLE. In conclusion, ADC values can be used for the diagnosis of invasive and DCIS as well as benign tumors. The NMLE lesions tend to have higher ADC values than mass lesions; therefore, the morphological appearance of a lesion needs to be considered when using the ADC value for diagnosis.

BTG2 inhibits the proliferation, invasion, and apoptosis of MDA-MB-231 triple-negative breast cancer cells.

The purposes of this study were to investigate the effects of B cell translocation gene 2 (BTG2) on the proliferation, apoptosis, and invasion of triple-negative breast cancer and to provide an experimental basis for the future treatment of human triple-negative breast cancer. A pcDNA3.1-BTG2 eukaryotic expression vector was constructed and transfected into the MDA-MB-231 human triple-negative breast cancer cell line using lipofection. Then, relevant changes in the biological characteristics of the BTG2-expressing cell line were analyzed using MTT (tetrazolium blue), flow cytometry, and Transwell invasion chamber assays. Additionally, the effects of BTG2 expression on cyclin D1, caspase 3, and matrix metalloproteinases 1/2 (MMP-1/-2) expression were analyzed. Cell proliferation was significantly lower in the pcDNA3.1-BTG2-transfected group compared to the empty vector and blank control groups (p<0.05). There was no significant difference between the empty vector and blank control groups. FCM results demonstrated that there were significantly more cells in the G1 phase of the cell cycle and fewer S phase cells in the pcDNA3.1-BTG2 group than in the empty vector and blank control groups (p<0.05). Additionally, the proportion of cells that migrated across the membrane was significantly lower in the pcDNA3.1-BTG2 group than in the empty vector and blank control groups (p<0.05). Cyclin D1 and MMP-1/-2 expression were significantly lower in MDA-MB-231 cells transfected with pcDNA3.1-BTG2 as compared to the empty vector and blank control groups (p<0.05). Caspase 3 expression was significantly higher in MDA-MB-231 cells from the pcDNA3.1-BTG2 group compared to the empty vector and blank control groups (p<0.05). In conclusion, BTG2 may inhibit MDA-MB-231 proliferation and promote apoptosis. Additionally, BTG2 may also inhibit the invasion of MDA-MB-231 human triple-negative breast cancer cells.

Expression and regulatory function of miRNA-182 in triple-negative breast cancer cells through its targeting of profilin 1.

We aimed to evaluate the expression of microRNA-182 (miR-182) in triple-negative breast cancer (TNBC) tissues and the TNBC cell line MDA-MB-231 and to investigate the effects of mirR-182 on the cellular behavior of MDA-MB-231 and the expression of the target gene profilin 1 (PFN1), thus providing new methods and new strategies for the treatment of TNBC. Quantitative real-time PCR (qRT-PCR) was utilized to evaluate the expression of miR-182 in TNBC tissues, relatively normal tissues adjacent to TNBC and the TNBC cell line MDA-MB-231. Forty-eight hours after the MDA-MB-231 cells were transfected with the miR-182 inhibitor, qRT-PCR was utilized to detect the changes in miR-182 expression levels, and an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was utilized to determine the effects of miR-182 on cell viability. Flow cytometry was adopted to determine whether miR-182 affects the proliferation rates and apoptosis levels of the MDA-MB-231 cells. The transwell migration assay method was used to investigate the effects of miR-182 on the migration of the MDA-MB-231 cells. A luciferase reporter gene system was applied to validate that PFN1 was the target gene of miR-182. Western blot was used to measure the effects of miR-182 on the PFN1 protein expression levels in the cells. qRT-PCR results showed that compared with the relatively normal tissues adjacent to TNBC, miR-182 expression was significantly increased in the TNBC tissues and the MDA-MB-231 cells (p<0.01). Compared with the control group, MDA-MB-231 cells transfected with the miR-182 inhibitor and incubated for 48 h showed significantly decreased miR-182 expression (p<0.01). The results of an MTT assay showed that inhibition of miR-182 in MDA-MB-231 cells led to significantly reduced cell viability (p<0.05). Flow cytometry analysis indicated that inhibition of miR-182 expression resulted in significantly decreased cell proliferation (p<0.05) and significantly increased levels of apoptosis (p<0.05). The results of a transwell migration assay showed that after inhibited of miR-182 expression, the number of cells passing through the transwell membranes was significantly decreased (p<0.05). The results from a luciferase reporter gene system showed that compared with the control group, the relative luciferase activity of the group transfected with the miR-182 inhibitor was significantly increased (p<0.05). Western blot analysis showed that compared with the control group, PFN1 protein expression levels were significantly increased in the MDA-MB-231 cells transfected with the miR-182 inhibitor and incubated for 48 h (p<0.05). In conclusion, miR-182 is upregulated in TNBC tissues and cells. It promotes the proliferation and invasion of MDA-MB-231 cells and could negatively regulate PFN1 protein expression. Treatment strategies utilizing inhibition of miR-182 expression or overexpression of the PFN1 gene might benefit patients with TNBC.

[Effects of mammalian-target-of-rapamycin pathway on lapatinib resistance in breast cancer MDA-MB-231 cells].

OBJECTIVE: To establish a lapatinib resistance cell line for elucidating the mechanisms of drug resistance of lapatinib in human breast cancer cells. METHODS: The human breast cancer MDA-MB-231 cells were exposed in an incremental dose of lapatinib to establish a lapatinib resistance rMDA-MB-231 cell line. The assay of methyl thiazolyl tetrazolium (MTT) was used to detect the cytotoxic activity of lapatinib against MDA-MB-231 and rMDA-MB-231 cells. The protein expression was detected by Western blot. Small interfering RNA was used to specifically knock down mammalian-target-of-rapamycin (mTOR) in rMDA-MB-231 cells. Apoptosis was determined by fluorescein isothiocyanate (FITC)-annexin V/PI staining and flow cytometry. RESULTS: The human breast cancer lapatinib resistance cell line rMDA-MB-231 was induced by lapatinib. The half maximal inhibitory concentration (IC50) values of lapatinib against MDA-MB-231 and rMDA-MB-231 cells were (6.1 ± 0.6) and (34.9 ± 2.7) µmol/L respectively (P < 0.01). Compared with MDA-MB-231 cells, the protein expression of mTOR in rMDA-MB-231 cells was significantly up-regulated. The protein expression of mTOR was significantly down-regulated by specific siRNA duplexes in rMDA-MB-231 cells. After siRNA interference, 20 µmol/L lapatinib was added into control, negative siRNA control and mTOR-targeted siRNA groups respectively. The percents of cell apoptosis in control, negative control and targeted siRNA groups were 13.4% ± 2.5%, 14.2% ± 2.8% and 34.6% ± 5.8% respectively, there was no significance between the first two groups (P > 0.05) , and there was significant difference between the control and targeted siRNA group (P < 0.01) . CONCLUSIONS: The up-regulation of mTOR plays an important role in the lapatinib-resistant phenotype of human breast cancer rMDA-MB-231 cells. And the down-regulation of mTOR increases the apoptotic death of lapatinib against rMDA-MB-231 cells.

Research progress and clinical evaluation of histotripsy: a narrative review.

Background and Objective: As a soft-tissue noninvasive ablation technology, high-intensity focused ultrasound (HIFU) has been widely used to treat many clinical diseases. However, traditional HIFU, based on thermal effects, has a high local working temperature, which may cause thermal damage to surrounding tissues and reduce the therapeutic effect. Based on the cavitation effect of HIFU, histotripsy can mechanically destroy the cells in the target lesion. This paper aims to explain the mechanism of histotripsy, summarize the research progress of animal models for clinical evaluation and clinical application, and analyze the advantages and limitations of histotripsy. Methods: Literature published from January 2006 to March 2022 was retrieved from the PubMed database. We reviewed these articles to examine histotripsy from the aspects of the mechanism, animal experiments, clinical trials, advantages, disadvantages, and optimization. Key Content and Findings: Histotripsy is a noninvasive, nonionizing, nonthermal ablation technique. The clinical application of histotripsy has made significant progress in the treatment of liver tumors, benign prostatic hyperplasia, and aortic valve calcification stenosis. Phase I clinical trials have demonstrated the safety and efficacy of histotripsy in the treatment of these diseases. More research is needed to evaluate and optimize its efficacy and safety and to fully explore its mechanism of action, pathological and immunological effects, and the short-term and long-term reactions of the body after treatment. Conclusions: Histotripsy has broad application prospects in ablation therapy and will benefit patients after more clinical trials are conducted in the future.

Neoadjuvant therapy with vs. without anthracyclines for HER2-positive breast cancer: a systematic review and meta-analysis.

Background: Neoadjuvant therapy has become the standard treatment for early human epidermal growth factor receptor 2 (HER2)-positive breast cancer, with most regimens using a combination of anti-HER2-targeted drugs and chemotherapy. However, the combination of anthracyclines and trastuzumab has high cardiac toxicity, and the efficacy evaluation of targeted therapy with or without anthracyclines is not unified. The purpose of this meta-analysis was to evaluate the relative efficacy and safety of anti-HER2-targeted therapy combined with vs. without anthracyclines neoadjuvant treatment. Methods: The following databases: PubMed, Medline, Embase, and Cochrane Library were systematically searched. Study inclusion was determined according to PICOS principles. PICOS: Patients, HER2-positive breast cancer; Intervention, anti-HER2-targeted therapy combined with anthracyclines; Control, without anthracyclines; Outcomes, the percentage of pathologic complete response (pCR), breast-conserving surgery (BCS), and grade 3 or worse adverse events according to CTCAE version 4.03; Studies, randomized controlled trials (RCTs) and retrospective studies. The meta-analysis was performed using RevMan5.3 software, and the odds ratio (OR) with 95% confidence intervals (CIs) was performed. Results: In total, 11 articles involving 1,998 patients were included with 1,155 patients in the anthracycline-containing group and 843 patients in the anthracycline-free group. For efficacy, there was no statistically significant difference in the percentage of pCR (OR 0.95; 95% CI: 0.61-1.48; P=0.83) and BCS (OR 1.18; 95% CI: 0.93-1.49; P=0.17) on anthracycline-free regimens compared with anthracycline-containing regimens. For safety, the combined effect values showed a significantly lower incidence of left ventricular ejection fraction decreases with the anthracycline-free regimen than with the anthracycline-containing regimen (OR 0.50; 95% CI: 0.35-0.71; P=0.0001). Other adverse effects and survival events were generally not statistically different in incidence between the two groups. The subgroup analysis suggested that hormone receptor status might be the source of heterogeneity in this study. Conclusions: Our study demonstrated that the targeted therapy combined with anthracyclines was associated with an increased risk of cardiac adverse events compared with the anthracycline-free group, with no significant difference in the percentage of pCR and BCS. Due to the high heterogeneity of this meta-analysis, more studies with longer follow-up are needed to validate the current findings and to further explore the removal and retention of anthracyclines.

Organising pneumonia caused by hormone (tamoxifen) therapy after radiotherapy for breast cancer: a case report and review of the literature.

Background: Five-year treatment with tamoxifen (TAM) has been the traditional standard of care for breast cancer. Organising pneumonia (OP) is a rare but significant complication of radiation therapy for breast cancer. The effect of TAM leading to OP has not yet been clearly documented. Case Description: This report describes the case of a 38-year-old female who developed progressive aggravation of round-like patchy bilateral pulmonary infiltrated with a reverse halo sign but without any clinical symptoms 5 months after TAM therapy, following breast-conserving surgery and radiotherapy (RT) for breast carcinoma. A lung biopsy was performed and revealed a histological pattern of OP. TAM therapy was discontinued, and subsequent gradual radiological improvement was observed. As there was no proof for TAM had caused the incident, TAM was re-administrated. Eight months after reinstitution of TAM, the same patchy migratory bilateral pulmonary infiltrated with reverse halo sign was found on chest CT with the patient claiming no discomforts nor any clinical symptoms. The diagnosis of TAM-related OP was made based on the exclusion of other causes and recurrence with the re-administration of TAM. The multidisciplinary team (MDT) concluded that TAM should be withdrawn and a "wait-and-see" approach was taken after a comprehensive assessment, instead of altering the medication or performing prophylactic mastectomy. Conclusions: The withdrawal and rechallenge of TAM strongly suggest that it may play a role as a cofactor in the occurrence of OP after RT for breast cancer, and RT may also be a cofactor in the occurrence of OP. It is extremely important to be alerted to the possibility of OP after concurrent or sequential hormonal therapy and RT.