PDF(Pubmed)
Abstract：
UNASSIGNED: Triple-negative breast cancer (TNBC) is characterized by its aggressive nature and absence of specific therapeutic targets, necessitating the reliance on chemotherapy as the primary treatment modality. However, the drug resistance poses a significant challenge in the management of TNBC. In this study, we investigated the role of DDX58 (DExD/H-box helicase 58), also known as RIG-I, in TNBC chemoresistance.
UNASSIGNED: The relationship between DDX58 expression and breast cancer prognosis was investigated by online clinical databases and confirmed by immunohistochemistry analysis. DDX58 was knockout by CRISPR-Cas9 system (DDX58-KO), knockdown by DDX58-siRNA (DDX58-KD), and stably over expressed (DDX58-OE) by lentivirus. Western blotting, immunofluorescence and qPCR were used for related molecules detection. Apoptosis was analyzed through flow cytometry (Annexin V/7AAD apoptosis assay) and Caspase 3/7 activity assay.
UNASSIGNED: Patients with lower expression of DDX58 led to lower rate of pathological complete response (pCR) and worse prognosis by online databases and hospital clinical data. DDX58-KD cells showed multiple chemo-drugs resistance (paclitaxel, doxorubicin, 5-fluorouracil) in TNBC cell lines. Similarly, DDX58-KO cells also showed multiple chemo-drugs resistance in a dosage-dependent manner. In the CDX model, tumours in the DDX58-KO group had a 25% reduction in the tumour growth inhibition rate (IR) compared to wild-type (WT) group after doxorubicin (Dox) treatment. The depletion of DDX58 inhibited proliferation and promoted the migration and invasion in MDA-MB-231 cells. The findings of our research indicated that DDX58-KO cells exhibit a reduction in Dox-induced apoptosis both in vivo and in vitro. Mechanistically, Dox treatment leads to a significant increase in the expression of double-stranded RNAs (dsRNAs) and activates the DDX58-Type I interferon (IFN) signaling pathway, ultimately promoting apoptosis in TNBC cells.
UNASSIGNED: In the process of TNBC chemotherapy, the deficiency of DDX58 can inhibit Dox-induced apoptosis, revealing a new pathway of chemotherapy resistance, and providing a possibility for developing personalized treatment strategies based on DDX58 expression levels.
UNASSIGNED: The relationship between DDX58 expression and breast cancer prognosis was investigated by online clinical databases and confirmed by immunohistochemistry analysis. DDX58 was knockout by CRISPR-Cas9 system (DDX58-KO), knockdown by DDX58-siRNA (DDX58-KD), and stably over expressed (DDX58-OE) by lentivirus. Western blotting, immunofluorescence and qPCR were used for related molecules detection. Apoptosis was analyzed through flow cytometry (Annexin V/7AAD apoptosis assay) and Caspase 3/7 activity assay.
UNASSIGNED: Patients with lower expression of DDX58 led to lower rate of pathological complete response (pCR) and worse prognosis by online databases and hospital clinical data. DDX58-KD cells showed multiple chemo-drugs resistance (paclitaxel, doxorubicin, 5-fluorouracil) in TNBC cell lines. Similarly, DDX58-KO cells also showed multiple chemo-drugs resistance in a dosage-dependent manner. In the CDX model, tumours in the DDX58-KO group had a 25% reduction in the tumour growth inhibition rate (IR) compared to wild-type (WT) group after doxorubicin (Dox) treatment. The depletion of DDX58 inhibited proliferation and promoted the migration and invasion in MDA-MB-231 cells. The findings of our research indicated that DDX58-KO cells exhibit a reduction in Dox-induced apoptosis both in vivo and in vitro. Mechanistically, Dox treatment leads to a significant increase in the expression of double-stranded RNAs (dsRNAs) and activates the DDX58-Type I interferon (IFN) signaling pathway, ultimately promoting apoptosis in TNBC cells.
UNASSIGNED: In the process of TNBC chemotherapy, the deficiency of DDX58 can inhibit Dox-induced apoptosis, revealing a new pathway of chemotherapy resistance, and providing a possibility for developing personalized treatment strategies based on DDX58 expression levels.
摘要：
■三阴性乳腺癌（TNBC）的特征是其侵袭性和缺乏特定的治疗靶标，需要依赖化疗作为主要治疗方式。然而,耐药性对TNBC的管理提出了重大挑战。在这项研究中,我们研究了DDX58（DExD/H-box解旋酶58）的作用，也被称为RIG-I,在TNBC化学抗性中。
■通过在线临床数据库研究DDX58表达与乳腺癌预后之间的关系，并通过免疫组织化学分析证实。DDX58被CRISPR-Cas9系统(DDX58-KO)敲除,DDX58-siRNA（DDX58-KD）并且通过慢病毒稳定过表达(DDX58-OE)。西方印迹,免疫荧光和qPCR用于相关分子检测。通过流式细胞术(膜联蛋白V/7AAD细胞凋亡测定)和Caspase3/7活性测定分析细胞凋亡。
■根据在线数据库和医院临床数据，DDX58表达较低的患者病理完全缓解率（pCR）较低，预后较差。DDX58-KD细胞显示多种化疗药物耐药(紫杉醇，阿霉素,5-氟尿嘧啶）在TNBC细胞系中。同样，DDX58-KO细胞也以剂量依赖性方式显示多种化疗药物抗性。在CDX模型中，与多柔比星（Dox）治疗后的野生型（WT）组相比，DDX58-KO组的肿瘤生长抑制率（IR）降低了25％。DDX58的清除抑制MDA-MB-231细胞的增殖，促进细胞的迁移和侵袭。我们的研究结果表明，DDX58-KO细胞在体内和体外均表现出Dox诱导的凋亡减少。机械上,Dox处理导致双链RNA（dsRNA）的表达显着增加，并激活DDX58-I型干扰素（IFN）信号通路，最终促进TNBC细胞凋亡。
■在TNBC化疗过程中，DDX58的缺乏可以抑制Dox诱导的细胞凋亡，揭示了化疗耐药的新途径，并为开发基于DDX58表达水平的个性化治疗策略提供了可能性。
■通过在线临床数据库研究DDX58表达与乳腺癌预后之间的关系，并通过免疫组织化学分析证实。DDX58被CRISPR-Cas9系统(DDX58-KO)敲除,DDX58-siRNA（DDX58-KD）并且通过慢病毒稳定过表达(DDX58-OE)。西方印迹,免疫荧光和qPCR用于相关分子检测。通过流式细胞术(膜联蛋白V/7AAD细胞凋亡测定)和Caspase3/7活性测定分析细胞凋亡。
■根据在线数据库和医院临床数据，DDX58表达较低的患者病理完全缓解率（pCR）较低，预后较差。DDX58-KD细胞显示多种化疗药物耐药(紫杉醇，阿霉素,5-氟尿嘧啶）在TNBC细胞系中。同样，DDX58-KO细胞也以剂量依赖性方式显示多种化疗药物抗性。在CDX模型中，与多柔比星（Dox）治疗后的野生型（WT）组相比，DDX58-KO组的肿瘤生长抑制率（IR）降低了25％。DDX58的清除抑制MDA-MB-231细胞的增殖，促进细胞的迁移和侵袭。我们的研究结果表明，DDX58-KO细胞在体内和体外均表现出Dox诱导的凋亡减少。机械上,Dox处理导致双链RNA（dsRNA）的表达显着增加，并激活DDX58-I型干扰素（IFN）信号通路，最终促进TNBC细胞凋亡。
■在TNBC化疗过程中，DDX58的缺乏可以抑制Dox诱导的细胞凋亡，揭示了化疗耐药的新途径，并为开发基于DDX58表达水平的个性化治疗策略提供了可能性。
