RESUMEN
Triple-negative breast cancer (TNBC) is characterized by high mortality rates primarily due to its propensity for metastasis. Addressing this challenge necessitates the development of effective antimetastatic therapies. This study aimed to identify natural compounds with potential antimetastatic properties mainly based on the high-throughput phenotypic screening system. This system, utilizing luciferase reporter gene assays combined with scratch wound assays, evaluates compounds based on their influence on the epithelial-mesenchymal transition (EMT) marker E-cadherin. Through this approach, aurovertin B (AVB) was revealed to have significant antimetastatic capability. Notably, AVB exhibited substantial metastasis suppression in many TNBC cell lines, including MDA-MB-231, HCC1937 and 4T1. Also, its remarkable antimetastatic activity was demonstrated in vivo via the orthotopic breast cancer mouse model. Further exploration revealed a pronounced association between AVB-induced upregulation of DUSP1 (dual-specificity phosphatase 1) and its inhibitory effect on TNBC metastasis. Additionally, microarray analysis conducted to elucidate the underlying mechanism of the AVB-DUSP1 interaction identified ATF3 (activating transcription factor 3) as a critical transcription factor instrumental in DUSP1 transcriptional activation. This discovery, coupled with observations of enhanced ATF3-DUSP1 expression and consequent reduction in TNBC metastatic foci in response to AVB, provides novel insights into the molecular mechanisms driving metastasis in TNBC. Significance Statement We construct a high-throughput phenotypic screening system utilizing EMT marker E-cadherin promoter luciferase reporter gene combined with scratch wound assays. Aurovertin B was revealed to possess significant antimetastatic activity through this approach, which was further demonstrated via in vivo and in vitro experiments. The discovery of the regulatory role of the ATF3-DUSP1 pathway enriches our understanding of TNBC metastasis mechanism and suggests the potential of ATF3 and DUSP1 as biomarkers for diagnosing TNBC metastasis.
RESUMEN
In order to obtain the lead compound for treatment of rheumatoid arthritis (RA), in this study, therapeutic efficacy of three bispecific antibodies (BsAB-1, BsAB-2 and BsAB-3) against both hIL-1beta and hIL-17 were compared on CIA model mice. First, by ELISA method we compared the binding capacity of the three bispecific antibodies to the two antigens. The results showed that all three antibodies could simultaneously bind both antigens, among these antibodies, BsAB-1 was superior over BsAB-2 and BsAB-3. CIA model was established with chicken type II collagen (CII) and developed RA-like symptoms such as ankle swelling, skin tight, hind foot skin hyperemia. The CIA mice were treated with three antibodies once every two days for total of 29 days. Compared with the CIA model mice, the RA-like symptoms of the antibody treated-mice significantly relieved, while the BsAB-1 treated-mice were almost recovered. CII antibody level in the serum and cytokines (IL-2, IL-1beta, IL-17A and TNF-alpha) expression in the spleen were examined. Compared with the CIA model mice, all three antibodies could significantly reduce CII antibody and cytokine expression levels. BsAB-1 antibody was more potent than BsAB-2 and BsAB-3. In summary, BsAB-1 is superior over BsAB-2 and BsAB-3 in amelioration of RA symptoms and regulation of CII antibody production and pro-inflammatory cytokine expression, therefore, BsAB-1 can be chosen as a lead compound for further development of drug candidate for treatment of RA.
Asunto(s)
Anticuerpos Biespecíficos/uso terapéutico , Artritis Experimental/terapia , Artritis Reumatoide/terapia , Animales , Anticuerpos/metabolismo , Anticuerpos Biespecíficos/inmunología , Reacciones Antígeno-Anticuerpo , Artritis Experimental/inducido químicamente , Artritis Experimental/metabolismo , Artritis Reumatoide/inducido químicamente , Artritis Reumatoide/metabolismo , Colágeno Tipo II/inmunología , Interleucina-17/metabolismo , Interleucina-1beta/metabolismo , Interleucina-2/metabolismo , Masculino , Ratones , Bazo/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
CRISPR/Cas genome editing is a simple, cost effective, and highly specific technique for introducing genetic variations. In mammalian cells, CRISPR/Cas can facilitate non-homologous end joining, homology- directed repair, and single-base exchanges. Cas9/Cas12a nuclease, dCas9 transcriptional regulators, base editors, PRIME editors and RNA editing tools are widely used in basic research. Currently, a variety of CRISPR/Cas-based therapeutics are being investigated in clinical trials. Among many new findings that have advanced the field, we highlight a few recent advances that are relevant to CRISPR/Cas-based gene therapies for monogenic human genetic diseases.
Asunto(s)
Sistemas CRISPR-Cas , Edición Génica , Enfermedades Genéticas Congénitas/terapia , Terapia Genética , Animales , Línea Celular , HumanosRESUMEN
<p><b>OBJECTIVE</b>To explore the effectiveness and safety of laparoscopic presacral neurectomy (LPN) in treating endometriosis-associated pain.</p><p><b>METHODS</b>Totally 64 patients with endometriosis were divided into two groups using prospective non-random method. Patients in the control group received only the conventional laparoscopic resection of endometriosis lesions, while patients in the LPN group underwent LPN in addition to the resection of endometriosis lesions. The pre-operative pain scores, intra-operative staging results, surgical duration, intra-surgical blood loss, post-operative pain relief were compared between these two groups.</p><p><b>RESULTS</b>These two groups showed no significant differences in terms of age, body weight, pre-operative pain score, surgery staging, surgical duration, and intra-operative blood loss (all P > 0.05). All patients were followed up for 6 to 18 months (median: 12.8 months). The post-operative pain relief rate was 89.28% (25/ 28) in LPN group and 61.29% (19/31) in the control group (P = 0.030).</p><p><b>CONCLUSION</b>LPN can effectively and safely in treating endometriosis and its associated pain.</p>