Background/Aim
Berberine (BBR) is known to be effective at inhibiting cell proliferation and promoting apoptosis in various cancer cells. However, the effects of BBR on triple‐negative breast cancer ...(TNBC) cells remain unclear. The aim of this study was to investigate the cell inhibition effects of BBR on different subtypes of TNBC cells.
Methods
Using human TNBC cell lines of different subtypes, namely, MDA‐MB‐231, MDA‐MB‐468, MDA‐MB‐453, and BT‐549 as in vitro models, antiproliferative effects of BBR were investigated by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay, trypan blue exclusion assay, and clonogenic assay. Furthermore, cell apoptosis and autophagy were analyzed by flow cytometry, immunofluorescent staining, and LC3 I/II‐targeted Western blotting. Various cell growth‐related signaling pathways (AKT/ERK/p38) and the expression of proteins present in various cell cycle kinase complexes were analyzed by Western blotting.
Results
BBR concentration‐dependently suppressed cell proliferation in MDA‐MB‐468 (0, 3, 6, and 12 μM) and MDA‐MB‐231 (0, 6.25, 12.5, and 25 μM). The inhibitory effect was not brought about by inducing cell apoptosis, necrosis, or autophagy. Cell cycle analysis disclosed an increased S+G2/M fraction among the BBR‐treated MDA‐MB‐231 and MDA‐MB‐453 cells; while with the BBR‐treated MDA‐MB‐468 and BT‐549 lines, an increased G0/G1 fraction was found. In MDA‐MB‐231 and MDA‐MB‐453 cells, by Western blotting, BBR decreased the expression of Cyclin A and CDK1, On the other hand, in BBR‐treated MDA‐MB‐468 and BT‐549 cells, there was a decrease in Cyclin D and CDK4 expression.
Conclusion
Our results demonstrate that the antiproliferation effects of BBR occur via different mechanisms in different subtypes of TNBC cells, which suggests that BBR has potential as a personalized treatment for TNBC patients.
Our results demonstrate that the antiproliferation effects of berberine (BBR) occur via different mechanisms in the four representative triple‐negative breast cancer (TNBC) cell lines, which suggests that BBR has potential as a personalized treatment for TNBC patients
How Does Moxibustion Possibly Work? Chiu, Jen-Hwey
Evidence-based complementary and alternative medicine,
01/2013, Volume:
2013
Journal Article
Peer reviewed
Open access
“Acupmoxa” is a hybrid word of “acupuncture” and “moxibustion” that more closely resembles the Chinese ideograph for this treatment. People in Western countries are more familiar with acupuncture, ...while moxibustion is less popular, partially due to the paucity of scientific studies. Although the evidence-based efficacy of moxibustion needs to be further clarified, the mechanisms by which moxibustion may work include temperature-related and nontemperature-related ones. Local somatothermal stimulation (LSTS), one type of moxibustion, is achieved by application of a heat source to and above the acupoint. Such mild heat stimulation of the acupoint induces little skin damage, in contrast to the burning effect of moxibustion, but does provoke mild oxidative stress in the viscera. Thus, preconditioned LSTS at the peripheral acupoints LR 14 and PC 6 of animals is able to induce visceral HSP70 expression and to protect the liver and the heart against ischemia-reperfusion injury. Nontemperature-related mechanisms include smoke, herbs, and biophysical (far infrared) stimulation. We conclude that LSTS, a remote preconditioning method, has potential clinical usefulness. However, evidence-based efficacy and safety studies involving large-scaled clinical trials are needed in order that this approach will pass muster with Western scientists.
There is good evidence that the tumor microenvironment plays an important role in cancer metastasis and progression. Our previous studies have shown that brain-derived neurotrophic factor (BDNF) ...participates in the process of metastasis and in the migration of cancer cells. The aim of this study was to investigate the role of BDNF on the tumor cell microenvironment, namely, the cancer cell-endothelial cell interaction of TNBC cells.
We conducted oligoneucleotide microarray analysis of potential biomarkers that are able to differentiate recurrent TNBC from non-recurrent TNBC. The MDA-MB-231 and human endothelial HUVEC lines were used for this study and our approaches included functional studies, such as migration assay, as well as Western blot and real-time PCR analysis of migration and angiogenic signaling. In addition, we analyzed the survival outcome of TNBC breast cancer patients according to their expression level of BDNF using clinical samples.
The results demonstrated that BDNF was able to bring about autocrinal (MDA-MB-231) and paracrinal (HUVECs) regulation of BDNF-TrkB gene expression and this affected cell migratory activity. The BDNF-induced migratory activity was blocked by inhibitors of ERK, PI3K and TrkB when MDA-MB-231 cells were examined, but only an inhibitor of ERK blocked this activity when HUVEC cells were used. Furthermore, decreased migratory activity was found for △BDNF and △TrkB cell lines. Ingenuity pathway analysis (IPA) of MDA-MB-231 cells showed that BDNF is a key factor that is able to regulate a network made up of metalloproteases and calmodulin. Protein expression levels in a tissue array of tumor slices were found to be correlated with patient prognosis and the results showed that there was significant correlation of TrkB expression, but not of BDNF. expressionwith patient DFS and OS.
Our study demonstrates that up-regulation of the BDNF signaling pathway seems tobe involved in the mechanism associated with early recurrence in triple negative breast cancer cell. In addition, BDNF can function in either an autocrine or a paracrine manner to increase the migration ability of both MDA-MB-231 cells and HUVEC cells. Finally, overexpression of TrkB, but not of BDNF, is significantly associated with a poor survival outcome for TNBC patients.
Background
The aim of this study was to investigate the role of IL-17A in the cancer microenvironment and the recurrence of triple negative breast cancer
(
TNBC).
Methods
Using human TNBC cell lines, ...the role of IL17-A was investigated by knocked down of IL-17A (Δ
IL-17A
) and by administration of IL-17A into the culture medium. Cell proliferation assays, migration assays, as well as Western blot analysis and real-time PCR, were used to evaluate IL-17A-related signaling. Three types of 4T1 cells were implanted into BALB/c mice, namely wild type (WT), Δ
IL-17A
, and WT + neutralizing IL-17 antibody (WT + Ab) cells. Tumor weight, necrosis area, and the number of circulating tumor cells (CTCs) were measured. Immunohistochemistry and Western blotting were used to analyze expression of CD34, CD8, and TGF-β1 as well as anoikis resistance. The Kaplan–Meier’s method was used to correlate IL-17A expression and patient outcome, including disease-free survival (DFS) and overall survival (OS).
Results
Our results demonstrated that IL-17A was able to stimulate the migratory activity, but not the growth rate, of MDA-MB-231/468 cells. In vivo, for the ΔIL-17A group, there was an increase in necrosis area, a decrease in tumor CD34 expression and a reduction in the number of CTCs. Furthermore, in WT + Ab group, there was a decreased in tumor expression of CD34, fewer CD8 ( +) cells, and fewer CTCs, but an increase in expression of TGF-β1 expression. Both of the above were compared to the WT group. Knockdown of IL-17A also decreased anoikis resistance in human TNBC and the murine 4T1 cell lines. Kaplan–Meier analysis disclosed a negative correlation between tumor expression of IL-17A and OS in TNBC patients.
Conclusion
We conclude that IL-17A promotes migratory and angiogenic activity in tumors, enhances anoikis resistance, and modulates the immune landscape of the tumor microenvironment such changes favor cancer metastasis.
Purpose
Deleterious germline
BRCA1
/
2
mutations are among the most highly pathogenic variants in hereditary breast and ovarian cancer syndrome. Recently, genes implicated in homologous recombination ...repair (HRR) pathways have been investigated extensively. Defective HRR genes may indicate potential clinical benefits from PARP (poly ADP ribose polymerase) inhibitors beyond
BRCA1
/
2
mutations.
Methods
We evaluated the prevalence of
BRCA1
/
2
mutations as well as alterations in HRR genes with targeted sequencing. A total of 648 consecutive breast cancer samples were assayed, and HRR genes were evaluated for prevalence in breast cancer tissues.
Results
Among 648 breast cancers, there were 17 truncating and 2 missense mutations in
BRCA1
and 45 truncating and 1 missense mutation in
BRCA2
, impacting 3% and 5% of the study population (collectively altered in 6%) with cooccurrence of
BRCA1
/
2
in 7 breast cancers. On the other hand, HRR genes were altered in 122 (19%) breast cancers, while TBB (Talazoparib Beyond BRCA) trial-interrogated genes (excluding
BRCA1
/
2
) were mutated in 107 (17%) patients. Beyond
BRCA1/2
, the most prevalent HRR mutant genes came from
ARID1A
(7%),
PALB2
(7%), and
PTEN
(6%). Collectively, 164 (25%) of the 648 Taiwanese breast cancer samples harbored at least one mutation among HRR genes.
Conclusions
The prevalence of
BRCA1
/
2
mutations was far below one tenth, while the prevalence of HRR mutations was much higher and approached one-fourth among Taiwanese breast cancers. Further opportunities to take advantage of defective HRR genes for breast cancer treatment should be sought for the realization of precision medicine.
The homologous recombination (HR) repair pathway for DNA damage, particularly the BRCA1 and BRCA2 genes, has become a target for cancer therapy, with poly ADP-ribose polymerase (PARP) inhibitors ...showing significant outcomes in treating germline BRCA1/2 (gBRCA1/2) mutated breast cancer. Recent studies suggest that some patients with somatic BRCA1/2 (sBRCA1/2) mutation or mutations in HR-related genes other than BRCA1/2 may benefit from PARP inhibitors as well, particularly those with PALB2 mutations. The current analysis aims to evaluate the prevalence of genetic alterations specific to BRCA1, BRCA2, and PALB2 in a large cohort of Taiwanese breast cancer patients through tumor-targeted sequencing.
A total of 924 consecutive assays from 879 Taiwanese breast cancer patients underwent tumor-targeted sequencing (Thermo Fisher Oncomine Comprehensive Assay v3). We evaluated BRCA1, BRCA2, and PALB2 mutational profiles, with variants annotated and curated by the ClinVAR, the Oncomine™ Knowledgebase Reporter, and the OncoKB™. We also conducted reflex germline testing using either whole exome sequencing (WES) or whole genome sequencing (WGS), which is ongoing.
Among the 879 patients analyzed (924 assays), 130 had positive mutations in BRCA1 (3.1%), BRCA2 (8.6%), and PALB2 (5.2%), with a total of 14.8% having genetic alterations. Co-occurrence was noted between BRCA1/BRCA2, BRCA1/PALB2, and BRCA2/PALB2 mutations. In BRCA1-mutated samples, only p.K654fs was observed in three patients, while other variants were observed no more than twice. For BRCA2, p.N372H was the most common (26 patients), followed by p.S2186fs, p.V2466A, and p.X159_splice (5 times each). For PALB2, p.I887fs was the most common mutation (30 patients). This study identified 176 amino acid changes; 60.2% (106) were not documented in either ClinVAR or the Oncomine™ Knowledgebase Reporter. Using the OncoKB™ for annotation, 171 (97.2%) were found to have clinical implications. For the result of reflex germline testing, three variants (BRCA1 c.1969_1970del, BRCA1 c.3629_3630del, BRCA2 c.8755-1G > C) were annotated as Pathogenic/Likely pathogenic (P/LP) variants by ClinVar and as likely loss-of-function or likely oncogenic by OncoKB; while one variant (PALB2 c.448C > T) was not found in ClinVar but was annotated as likely loss-of-function or likely oncogenic by OncoKB.
Our study depicted the mutational patterns of BRCA1, BRCA2, and PALB2 in Taiwanese breast cancer patients through tumor-only sequencing. This highlights the growing importance of BRCA1/2 and PALB2 alterations in breast cancer susceptibility risk and the treatment of index patients. We also emphasized the need to meticulously annotate variants in cancer-driver genes as well as actionable mutations across multiple databases.
Aim
To determine whether acupressure can prevent or relieve the adverse drug reactions (ADRs) of anti‐tuberculosis drugs.
Background
People receiving drug treatment for TB often experience ADRs that ...may cause them to stop taking their medication. Acupressure is a form of traditional Chinese medicine that can be applied to alleviate or prevent disease symptoms.
Design
A double‐blinded, repeated‐measures clinical trial in hospitals in Taiwan was carried out from April 2015 – May 2017.
Methods
Convenience sampling was used to select 32 people (15 for the experimental group and 17 for the control group) aged >20 years who were taking anti‐tuberculosis drugs. The people were randomized to receive 4‐week of true acupressure and 4‐weeks of sham acupressure. Acupressure therapy was given by a researcher in all cases. Both groups received treatment once per day on weekdays, with 15 min for each acupressure session. Outcomes (gastrointestinal irritation and adverse skin reactions) were assessed according to the people feedback and the physicians’ recordings during the treatment course, and during monthly follow‐up visits for 6 months thereafter.
Results
Both groups typically experienced gastrointestinal irritation and adverse skin reactions within 2 months of beginning anti‐tuberculosis drug treatment. The 4‐weeks intervention involving relevant acupressure points successfully relieved both types of side effects in both immediate and delayed manner.
Conclusions
When correctly implemented, acupressure can prevent and relieve the ADRs of anti‐tuberculosis drugs, and motivate people to complete their treatment course.
目的
确定穴位按压能否预防或减轻抗结核药物的不良反应。
背景
接受结核病药物治疗的患者经常会出现不良反应,他们可能会因此停药。穴位按压是一种中医治疗方法,可以用来缓解或预防疾病症状。
设计
2015年4月至2017年5月,在台湾的医院进行了一项双盲、重复测量的临床试验。
方法
采用便利抽样原则,共选择了32名患者(实验组15名,对照组17名),患者年龄均大于20岁,并正在服用抗结核药物。随机分配患者接受4周的真实穴位按压或4周的假穴位按压。研究人员对所有患者进行了指压疗法。两组在工作日每天接受一次治疗,每次穴位按压疗程15分钟。治疗过程中,以及随后6个月的每月随访中,根据患者的反馈和医生的记录评估结果(胃肠道刺激和皮肤不良反应)。
结果
两组患者一般在开始抗结核药物疗法后的两个月内出现胃肠道刺激和皮肤不良反应。经过为期4周对相关穴位的按压干预后,这两种副作用分别即时和延迟缓解。
结论
如果实施正确,穴位按压可以预防和减轻抗结核药物疗法的不良反应,并鼓励患者完成的治疗过程。
Introduction
The aim of the study was to perform digital RNA counting to validate a gene expression signature for operable breast cancers initially treated with curative intention, and the risk of ...recurrence, distant metastasis, and mortality was predicted.
Methods
Candidate genes were initially discovered from the coherent genomic and transcriptional alternations from microarrays, and the extended concurrent genes were used to build a risk stratification model from archived formalin-fixed paraffin-embedded (FFPE) tissues with the NanoString nCounter.
Results
The extended concurrent genes signature was prognostic in 144 Taiwanese breast cancers (5-year relapse-free survival: 89.8 and 69.4% for low- and high-risk group, log-rank test:
P
= 0.004). Cross-platform comparability was evidenced from significant and positive correlations for most genes as well as equal covariance matrix across 64 patients assayed for both microarray and digital RNA counting.
Discussion
Archived FFPE samples could be successfully assayed by the NanoString nCounter. The purposed signature was prognostic stratifying breast cancer patients into groups with distinct survival patterns, and clinical applicability of the residual risk model was proved.
The aim of this study was to elucidate molecular profiling in HER2-low tumors based on a promising dataset. A total of 615 consecutive HER2-negative breast cancer samples were assayed. The genomic ...mutations in the two groups with different HER2 expression levels (HER2-0 vs. HER2-low) were compared. The mutation types obtained via next-generation targeted sequencing were correlated with the clinicopathological features of the patients with HER2-0 and HER2-low breast cancer. The results showed that there was a significantly higher percentage of receptor-positive (ER/PR) tumors and more low-level Ki-67 tumors, but a lower incidence of stage I/II tumors in the HER2-low group compared to the HER2-0 group. There was a significantly higher frequency of 17.62% (65/369) for
_SNA in the HER2-low group than in the HER2-0 group, which had a frequency of only 9.35% (23/246) (
= 0.006). When the called gene alterations in the triple-negative breast cancer (TNBC) group were compared with those in the luminal-like breast cancer group, there was a significantly high frequency of 28.17% (140/497) for
_SNA in a luminal-like group than in the TNBC group(16.95% (20/118)).We conclude that the early detection of
mutations is likely to be important and might help therapeutic decision making in patients with HER2-low tumors.
Aims
Invasive breast cancer patients with human epidermal growth factor receptor 2 (HER2) immunohistochemical (IHC) scores of 3+ or 2+ with reflex in‐situ hybridisation (ISH) positivity are suitable ...for anti‐HER2 therapies. The aim of this study is to investigate whether the prognoses between IHC 3+ patients and IHC 2+/ISH+ patients are different.
Methods and results
We analysed the clinicopathological information of 886 consecutive cases of HER2‐positive early breast cancer. The influences of the patients’ age, cancer stage, hormone receptor status and anti‐HER2 treatment were adjusted using a multivariate Cox regression model. Both HER2 copy numbers and HER2 ISH ratios of the IHC 3+ group were significantly higher than those of the IHC 2+/ISH+ group. The outcomes of IHC 3+ patients were significantly better than those of IHC 2+/ISH+ patients in the univariate and multivariate analyses. HER2 copy numbers of ≥8 represented the best prognostic value, and it was chosen to be the cut‐off value. The reflex ISH for IHC 2+ patients with high HER2 copy numbers (≥8) predicted a better overall survival than that for those with low HER2 copy numbers.
Conclusion
HER2 IHC scores and HER2 copy numbers can provide prognostic information for patients with HER2‐positive invasive breast cancer. Both IHC 3+ and IHC 2+ patients with high HER2 copy numbers had a better prognosis.
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