CN113952452A - 一种基于碳点的肿瘤放疗增敏剂及其应用 - Google Patents
一种基于碳点的肿瘤放疗增敏剂及其应用 Download PDFInfo
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Abstract
本发明涉及一种基于碳点的肿瘤放疗增敏剂及其应用，其特征在于：碳点具有较高的生物相容性，对生物各脏器均无损伤。其次碳点利用材料本身的EPR效应被动靶向肿瘤组织，使肿瘤细胞的细胞周期停滞于G2/M期，抑制肿瘤细胞的增殖。碳点消耗肿瘤部位的GSH并产生大量的ROS，促进DNA损伤诱导肿瘤细胞凋亡。肿瘤细胞处于G2/M期时，对放疗的敏感性较高。细胞内ROS水平的升高，可提高肿瘤细胞的固有辐射敏感性。基于以上两个特点，碳点可增强放射线对肿瘤细胞的敏感性，实现肿瘤放疗增敏目的。综上所述，碳点可实现靶向肿瘤细胞、抑制肿瘤细胞增殖、和增强放疗敏感性的多重效能，具有较好的应用前景。
Description
技术领域
本发明涉及碳纳米材料化学和生物化学的交叉领域，具体涉及一种用于肿瘤放疗增敏的碳点及其应用。
背景技术
恶性肿瘤（癌症）是威胁人类健康的重大疾病。目前放射治疗已被广泛应用于多种癌症的辅助治疗。然而，存在对放疗的耐药性、不良反应较多和治疗效果较差等问题。在恶性肿瘤放疗中可以通过抑制DNA损伤修复、调节细胞周期分布、诱导活性氧（ROS）的生成以及改变肿瘤细胞代谢等方面增强肿瘤细胞对射线的敏感性。现有的放疗增敏药物仅能部分改善放疗敏感性，且伴随着严重的不良反应，在临床上的应用具有一定的局限性。
目前，半导体材料等放疗增敏药物的研究与应用已有报道，但由于尺寸的限制，阻碍了肾脏的清除能力，长期积累导致肝脏损伤。荧光纳米碳点由于具有分子量小、荧光稳定性高、具有生物相容性好且毒性低等特点，目前已经开始替代传统的半导体材料用于肿瘤治疗领域。基于此，碳点在肿瘤放疗领域的应用成为可能。
发明内容
本发明设计了一种基于碳点的肿瘤放疗增敏剂及其应用，其解决的技术问题是现有增敏剂仅能部分改善放疗敏感性，且伴随着严重的不良反应，在临床上的应用具有一定的局限性。
为了解决上述存在的技术问题，本发明采用了以下方案：
一种基于碳点的肿瘤放疗增敏剂。
优选地，所述肿瘤包括鼻腔及鼻窦恶性肿瘤、鼻咽癌、口腔癌、喉癌、颅内肿瘤、甲状腺癌、舌癌、肺癌、食管癌、乳腺癌、胃癌、大肠癌、乙状结肠和直肠癌、肝癌、胰腺癌与壶腹周围癌、胆道癌、肾癌、前列腺癌、膀胱癌、睾丸恶性肿瘤、阴茎癌、子宫颈癌、子宫内膜癌、卵巢癌、纤维组织细胞癌、横纹肌肉癌、滑膜肉瘤、黑色素瘤、骨肉瘤、尤文氏肉瘤、淋巴瘤和多发性骨髓瘤等恶性肿瘤中的一种或几种。
优选地，所述碳点为粒径小于等于10纳米的碳点。
一种基于碳点的肿瘤放疗增敏剂的应用，所述碳点具有较高的生物相容性，对生物各脏器均无损伤；碳点抑制肿瘤细胞的细胞周期停滞于G2/M期，同时消耗肿瘤部位的GSH并产生大量的ROS，增加放射线对肿瘤细胞的敏感性。
优选地，所述碳点为粒径小于等于10纳米的碳点。
优选地，步骤1、利用流式细胞仪检测不同浓度碳点对细胞周期的影响；步骤2、按照步骤1确定的浓度范围，按计划照射剂量分别按不同肿瘤细胞数接种于六孔板中，将不同浓度的碳点溶液加入到接种数量一定的肿瘤细胞中培养，克隆形成实验检测细胞的放疗敏感性；建立荷瘤小鼠模型，按照所述步骤1确定的浓度范围，不同计量X-射线对荷瘤小鼠的肿瘤治疗效果；按照所述步骤1确定的浓度范围，检测GSH和ROS水平。
优选地，所述建立荷瘤小鼠模型中所述碳点作用荷瘤小鼠浓度为前期实验结果已确定的生物安全浓度，其对小鼠内脏无损伤，且能改善荷瘤小鼠的肝脏功能。
优选地，所述X-射线的射线辐射计量0-10Gy。
该基于碳点的肿瘤放疗增敏剂及其应用具有以下有益效果：
（1）本发明中使用碳点的肿瘤放疗增敏剂，碳点可代谢，无副作用，可改善肝脏功能，通过将细胞阻滞于G2/M期，消耗GSH，促进ROS的产生，进而增加肿瘤细胞对放疗的敏感性。
（2）本发明使用碳点的肿瘤放疗增敏剂在实验中促进肿瘤细胞凋亡，同时改善荷瘤小鼠肝脏功能，可有效实现肿瘤治疗增敏效果并改善小鼠肝脏功能。
（3）本发明碳点利用材料本身的EPR效应被动靶向肿瘤组织，使肿瘤细胞的细胞周期停滞于G2/M期，抑制肿瘤细胞的增殖。其次碳点消耗肿瘤部位的GSH并产生大量的ROS，促进DNA损伤诱导肿瘤细胞凋亡。肿瘤细胞处于G2/M期时，对放疗的敏感性较高。细胞内ROS水平的升高，可提高肿瘤细胞的固有辐射敏感性。基于以上两个特点，碳点可增强放射线对肿瘤细胞的敏感性，实现肿瘤放疗增敏目的。综上所述，碳点可实现靶向肿瘤细胞、抑制肿瘤细胞增殖、和增强放疗敏感性的多重效能，具有较好的应用前景。
附图说明
图1为本发明碳点存在对肿瘤Lovo细胞系的细胞周期影响示意图；
图2为本发明中碳点在荷瘤小鼠各种内脏器官影响的示意图；
图3 本发明中碳点存在对肿瘤细胞系GSH含量的研究示意图；
图4本发明中碳点存在对肿瘤细胞系ROS含量的研究示意图。
具体实施方式
下面结合图1至图4，对本发明做进一步说明：
实施例1：
如图1所示，可以使用二甲双胍作为制备碳点的原材料，但是不限于二甲双胍，还可以是其他各种材料。确定优选的碳点的作用浓度50 ng/µL和100 ng/µL，作用时间24h和48h，流式细胞仪，检测细胞周期。附图1为对照组，50ng/µL，100ng/µL处理48h后lovo细胞的细胞周期的分布情况。
得出结论：碳点调节细胞周期分布，将肿瘤细胞的细胞周期停滞在放疗敏感的G2/M期。
实施例2：
如图2所示，建立荷瘤小鼠模型，并注射碳点溶液，优选浓度为0，12.5mg/Kg，25mg/Kg。
连续注射7天后，心脏采血处死小鼠并解剖，取各组瘤体，内脏器官，血液等，检测相关指标。其中附图2中的（1）、（2）、（3）分别代表0 mg/Kg，12.5mg/Kg，25mg/Kg。肉眼可见，使用碳点溶液越高的内脏肿瘤组织体积在变小。
得出结论：碳点可以代谢出体外，并改善荷瘤小鼠肝脏功能。
实施例3：
如图3所示，取重量相同的0 mg/Kg，12.5mg/Kg，25mg/Kg三组肿瘤组织。
使用试剂盒检测对照组和实验组GSH含量。GSH可以转化为GSSG，12.5mg/Kg，25mg/Kg处理组GSH/GSSH与0 mg/Kg相比，其比值明显下降，表明碳点处理组GSH大部分均转化为GSSG。
得出结论：碳点消耗GSH。
实施例3：
如图4所示，取重量相同的0 mg/Kg，12.5mg/Kg，25mg/Kg三组肿瘤组织。
使用试剂盒检测ROS含量。12.5mg/Kg处理组ROS含量显著高于0mg/Kg对照组，25mg/Kg处理组ROS含量虽然没有12.5mg/Kg处理组含量高，但是仍然高于0mg/Kg对照组。
得出结论：碳点促进肿瘤细胞ROS的产生。
上面结合附图对本发明进行了示例性的描述，显然本发明的实现并不受上述方式的限制，只要采用了本发明的方法构思和技术方案进行的各种改进，或未经改进将本发明的构思和技术方案直接应用于其它场合的，均在本发明的保护范围内。
Claims (8)
1.一种基于碳点的肿瘤放疗增敏剂。
2.根据权利要求1所述的基于碳点的肿瘤放疗增敏剂，其特征在于：所述碳点粒径小于等于10纳米的碳点。
3.根据权利要求1所述的基于碳点的肿瘤放疗增敏剂，其特征在于：所述肿瘤包括鼻腔及鼻窦恶性肿瘤、鼻咽癌、口腔癌、喉癌、颅内肿瘤、甲状腺癌、舌癌、肺癌、食管癌、乳腺癌、胃癌、大肠癌、乙状结肠和直肠癌、肝癌、胰腺癌与壶腹周围癌、胆道癌、肾癌、前列腺癌、膀胱癌、睾丸恶性肿瘤、阴茎癌、子宫颈癌、子宫内膜癌、卵巢癌、纤维组织细胞癌、横纹肌肉癌、滑膜肉瘤、黑色素瘤、骨肉瘤、尤文氏肉瘤、淋巴瘤和多发性骨髓瘤等恶性肿瘤中的一种或几种。
4.一种基于碳点的肿瘤放疗增敏剂的应用，其特征在于：所述碳点具有较高的生物相容性，对生物各脏器均无损伤；所述碳点利用该纳米材料本身的EPR效应实现被动靶向肿瘤组织，碳点抑制肿瘤细胞的细胞周期停滞于G2/M期，同时消耗肿瘤部位的GSH并产生大量的ROS，增加放射线对肿瘤细胞的敏感性。
5.根据权利要求4所述的基于碳点的肿瘤放疗增敏剂的应用，其特征在于：所述碳点粒径小于等于10纳米的碳点。
6.根据权利要求4或5所述的基于碳点的肿瘤放疗增敏剂的应用，其特征在于：
步骤1、利用流式细胞仪检测不同浓度碳点对细胞周期的影响；
步骤2、按照步骤1确定的浓度范围，按计划照射剂量分别按不同肿瘤细胞数接种于六孔板中，将不同浓度的碳点溶液加入到接种数量一定的肿瘤细胞中培养，克隆形成实验检测细胞的放疗敏感性；
建立荷瘤小鼠模型，按照所述步骤1确定的浓度范围，不同计量X-射线对荷瘤小鼠的肿瘤治疗效果；
按照所述步骤1确定的浓度范围，检测GSH和ROS水平。
7.根据权利要求4所述的基于碳点的肿瘤放疗增敏剂的应用，其特征在于：所述建立荷瘤小鼠模型中所述碳点作用荷瘤小鼠浓度为前期实验结果已确定的生物安全浓度，其对小鼠内脏无损伤，且能改善荷瘤小鼠的肝脏功能。
8.根据权利要求4-7中任何一项所述的基于碳点的肿瘤放疗增敏剂的应用，其特征在于：所述X-射线的射线辐射计量0-10Gy。
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