Exploring the Role of TRIP-Brs in Human Breast Cancer: An Investigation of Expression, Clinicopathological Significance, and Prognosis

Abstract

TRIP-Brs, a group of transcription factors (TFs) that modulate several mechanisms in higher organisms. However, the novel paradigm to target TRIP-Brs in specific cancer remains to be deciphered. In particular, comprehensive analysis of TRIP-Brs in clinicopathological and patients’ prognosis, especially in breast cancer (BRCA), is being greatly ignored. Therefore, we explored the key roles of TRIP-Br expression, modulatory effects, mutations, immune infiltration, and prognosis in BRCA using multidimensional approaches. We found elevated levels of TRIP-Brs in numerous cancer tissues than normal. Higher expression of TRIP-Br-2/4/5 was shown to be positively associated with lower survival, tumor grade, and malignancy of patients with BRCA. Additionally, higher TRIP-Br-3/4 were also significantly linked with worse/short survival of BRCA patients. TRIP-Br-1/4/5 were significantly overexpressed and enhanced tumorigenesis in large-scale BRCA datasets. The mRNA levels of TRIP-Brs have been also correlated with tumor immune infiltrate in BRCA patients. In addition, TRIP-Brs synergistically play a pivotal role in central carbon metabolism, cancer-associated pathways, cell cycle, and thyroid hormone signaling, which evoke that TRIP-Brs may be a potential target for the therapy of BRCA. Thus, this investigation may lay a foundation for further research on TRIP-Br-mediated management of BRCA.

Anti-breast cancer action of carbonic anhydrase IX inhibitor 4-[4-(4-Benzo[1,3]dioxol-5-ylmethyl-piperazin-1-yl)-benzylidene-hydrazinocarbonyl]-benzenesulfonamide (BSM-0004): in vitro and in vivo studies

“…Following 30 min incubation at room temperature in a humidified chamber, colourimetric measurements were done using a multiwell microplate reader (LEDETECT96 LED based 8 channel Microplate Absorbance Reader). Efficacy of BSM-0004 and morphological analysis were also observed by optical microscope and captured cells morphology images in a different dose [51][52][53][54] .…”

“…The sub-population of cells in various cell cycle phase was investigated by staining with PI. The cell checkpoints were observed using BD FACSCanto TM system according to reported previously [51][52][53][54] . Following to the treatment with 10 and 20 mM (with or without 200 mM CoCl 2 ) concentrations of BSM-0004 for 24 h, approximately $1 Â 10 6 cells MCF7 cells were harvested and fixed with 70% ethanol at room temperature by gently vortexed and kept at 4 C for overnight.…”

“…For the protein estimation, MCF7 cells were cultured in DMEM complete medium with 10 and 20 mM concentration of BSM-0004 and DMSO (control group), respectively. Next day, cells were harvested using pellet downed by centrifugation at 1000 rpm (Hanil Scientific, Inc., Gimpo, South Korea) for two minutes at 4 C. Then, cells were rinsed with 1Â PBS and followed the instruction as reported previously [51][52][53][54] . Briefly, cells pellet was spin-down and lysed in the RIPA lysis buffer with DTT, NaVO3, and PI, respectively.…”

“…The full experimental plan and methods were designed and approved by the Sookmyung Women's University's Board of Review Committee, and the Department of Animal Facility, Yongsan-gu, Seoul, Republic of Korea. The efficacy of BSM-0004 against MCF7 induced tumour burden in nude mice was carried out as described previously [51][52][53][54] . Briefly, totally 15 mice were sub-grouped in control (n ¼ 6), treatment (n ¼ 6), and normal (n ¼ 3) mice group, respectively.…”

“…The efficacy of BSM-0004 and oncogenic expression of tumour marker Ki67 and hCA-IX were evaluated by immunohistochemistry of tumour samples as described in the previous study [51][52][53][54] . Briefly, tumour tissues were fixed with 4% paraformaldehyde then embedded with high grade of paraffin.…”