PDF(Pubmed)
Abstract：
OBJECTIVE: Platinum-based drugs are cytotoxic drugs commonly used in cancer treatment. They cause DNA damage, effects of which on chromatin and cellular responses are relatively well described. Yet, the nuclear stress responses related to RNA processing are incompletely known and may be relevant for the heterogeneity with which cancer cells respond to these drugs. Here, we determine the type and extent of nuclear stress responses of prostate cancer cells to clinically relevant platinum drugs.
METHODS: We study nucleolar and Cajal body (CB) responses to cisplatin, carboplatin, and oxaliplatin with immunofluorescence-based methods in prostate cancer cells. We utilize organelle-specific markers NPM, Fibrillarin, Coilin, and SMN1, and study CB-regulatory proteins FUS and TDP-43 using siRNA-mediated downregulation.
RESULTS: Different types of prostate cancer cells have different sensitivities to platinum drugs. With equally cytotoxic doses, cisplatin, and oxaliplatin induce prominent nucleolar and CB stress responses while the nuclear stress phenotypes to carboplatin are milder. We find that Coilin is a stress-specific marker for platinum drug response heterogeneity. We also find that CB-associated, stress-responsive RNA binding proteins FUS and TDP-43 control Coilin and CB biology in prostate cancer cells and, further, that TDP-43 is associated with stress-responsive CBs in prostate cancer cells.
CONCLUSIONS: Our findings provide insight into the heterologous responses of prostate cancer cells to different platinum drug treatments and indicate Coilin and TDP-43 as stress mediators in the varied outcomes. These results help understand cancer drug responses at a cellular level and have implications in tackling heterogeneity in cancer treatment outcomes.
METHODS: We study nucleolar and Cajal body (CB) responses to cisplatin, carboplatin, and oxaliplatin with immunofluorescence-based methods in prostate cancer cells. We utilize organelle-specific markers NPM, Fibrillarin, Coilin, and SMN1, and study CB-regulatory proteins FUS and TDP-43 using siRNA-mediated downregulation.
RESULTS: Different types of prostate cancer cells have different sensitivities to platinum drugs. With equally cytotoxic doses, cisplatin, and oxaliplatin induce prominent nucleolar and CB stress responses while the nuclear stress phenotypes to carboplatin are milder. We find that Coilin is a stress-specific marker for platinum drug response heterogeneity. We also find that CB-associated, stress-responsive RNA binding proteins FUS and TDP-43 control Coilin and CB biology in prostate cancer cells and, further, that TDP-43 is associated with stress-responsive CBs in prostate cancer cells.
CONCLUSIONS: Our findings provide insight into the heterologous responses of prostate cancer cells to different platinum drug treatments and indicate Coilin and TDP-43 as stress mediators in the varied outcomes. These results help understand cancer drug responses at a cellular level and have implications in tackling heterogeneity in cancer treatment outcomes.
摘要：
目的：铂类药物是癌症治疗中常用的细胞毒性药物。它们会导致DNA损伤,其对染色质和细胞反应的影响相对较好地描述。然而，与RNA加工相关的核应激反应尚不完全清楚,可能与癌细胞对这些药物的反应异质性有关.这里,我们确定前列腺癌细胞对临床相关铂类药物的核应激反应的类型和程度.
方法：我们研究了核仁和Cajal体（CB）对顺铂的反应，卡铂,和奥沙利铂在前列腺癌细胞中的免疫荧光方法。我们利用细胞器特异性标记NPM，纤维素,Coilin,和SMN1，并使用siRNA介导的下调研究CB调节蛋白FUS和TDP-43。
结果：不同类型的前列腺癌细胞对铂类药物的敏感性不同。同样的细胞毒性剂量，顺铂,和奥沙利铂诱导显著的核仁和CB应激反应，而卡铂的核胁迫表型较温和。我们发现Coilin是铂类药物反应异质性的应激特异性标志物。我们还发现CB相关，应激反应RNA结合蛋白FUS和TDP-43控制前列腺癌细胞中的Coilin和CB生物学，进一步,TDP-43与前列腺癌细胞中的应激反应性CB有关。
结论：我们的研究结果为前列腺癌细胞对不同铂类药物治疗的异源反应提供了见解，并表明Coilin和TDP-43在不同的结果中作为应激介质。这些结果有助于了解细胞水平的癌症药物反应，并对解决癌症治疗结果的异质性具有重要意义。
方法：我们研究了核仁和Cajal体（CB）对顺铂的反应，卡铂,和奥沙利铂在前列腺癌细胞中的免疫荧光方法。我们利用细胞器特异性标记NPM，纤维素,Coilin,和SMN1，并使用siRNA介导的下调研究CB调节蛋白FUS和TDP-43。
结果：不同类型的前列腺癌细胞对铂类药物的敏感性不同。同样的细胞毒性剂量，顺铂,和奥沙利铂诱导显著的核仁和CB应激反应，而卡铂的核胁迫表型较温和。我们发现Coilin是铂类药物反应异质性的应激特异性标志物。我们还发现CB相关，应激反应RNA结合蛋白FUS和TDP-43控制前列腺癌细胞中的Coilin和CB生物学，进一步,TDP-43与前列腺癌细胞中的应激反应性CB有关。
结论：我们的研究结果为前列腺癌细胞对不同铂类药物治疗的异源反应提供了见解，并表明Coilin和TDP-43在不同的结果中作为应激介质。这些结果有助于了解细胞水平的癌症药物反应，并对解决癌症治疗结果的异质性具有重要意义。
