CN109692178A - 川楝素在制备抗乳腺癌化疗增效药物中的应用 - Google Patents
川楝素在制备抗乳腺癌化疗增效药物中的应用 Download PDFInfo
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Abstract
本发明的名称为川楝素在制备抗乳腺癌化疗增效药物中的应用,技术领域为本发明涉及川楝素逆转乳腺癌细胞对阿霉素耐受的新用途。本发明中,我们首次发现川楝素能显著增强阿霉素耐药的乳腺癌肿瘤细胞株MCF‑7/ADM对阿霉素诱导凋亡的敏感性。免疫荧光法显示川楝素明显促进阿霉素在耐药细胞MCF‑7/ADM中的积累,尤其是在细胞核中。进一步研究发现,川楝素能明显降低P‑gp蛋白表达水平。动物实验也表明川楝素和阿霉素联用对肿瘤有很好的抑制作用。这些结果表明川楝素联用阿霉素,能有效增强阿霉素对耐受性乳腺癌肿瘤的治效果。临床发现许多恶性肿瘤细胞对阿霉素具有耐药性,因此川楝素有较好的开发价值和应用前景。
Description
一、技术领域
本发明涉及川楝素逆转乳腺癌细胞对阿霉素耐受的新用途
二、背景技术
在全球范围内,乳腺癌位居女性癌症发病之首,全球每年新发乳腺癌病例约167.1万,每年约52.2万死于乳腺癌,发达国家的乳腺癌发病相对欠发达国家更高。在我国,乳腺癌是最主要的恶性肿瘤之一。乳腺癌发病率位居城乡女性首位,是危害居民生命健康的最主要的恶性肿瘤之一,且我国居民的癌症发病率在近十几年总体呈现上升趋势。流行病学调查显示,全球范围内,其发病率不断上升,且呈年轻化趋势,已成为女性恶性肿瘤之首,严重威胁患者的生命健康。
乳腺癌的诱发受到多种因素的综合影响。1、遗传因素是乳腺癌较为明确的危险因素,乳腺癌家族史尤其是一级亲属乳腺癌家族史对乳腺癌发病的影响最为直接。2、绝经时间晚为乳腺癌发生的危险因素,月经周期短、月经不规则也会增加乳腺癌的危险。母乳喂养可以降低乳腺癌的发病率。3、女性的一些日常生活习惯也与乳腺癌的发生息息相关。长期夜班族、饮食无规律亦是乳腺癌的危险因素,不规律的生活会使机体免疫力下降,内分泌失调,导致乳腺癌发病率增加。
乳腺癌起源于乳腺各级别导管和腺泡上皮,由腺上皮增生到不典型增生而逐步发展为原位癌、早期浸润癌至浸润性癌。根据世界卫生组织乳腺肿瘤病理学可将乳腺癌分为乳腺导管癌、浸润性导管癌、小叶癌和三阴性乳腺癌四种。不同级别的导管发生的癌变,其组织类型常常不同。乳腺癌中95%以上是恶性上皮性肿瘤,乳腺肉瘤十分少见。在中国女性乳腺癌中,70%以上为浸润性导管癌,其他组织类型,如浸润性导管和小叶癌、浸润性小叶癌和浸润性小叶癌合并其他型癌等。如今,临床上治疗乳腺癌仍是以手术切除为主,放化疗和免疫疗法治疗为辅,而化疗又是辅助疗法中最常采用也是效果最好的一种方法。在术后进行的化疗称为辅助化疗,其作用主要是杀灭原发病灶切除后的残余瘤细胞。新辅助化疗即术前采用化疗,可以缩小乳腺癌原发肿瘤的体积,使部分肿瘤患者从最开始的不可切除转化为可以切除,使更多的乳腺癌患者可以进行保留乳房的手术,对于较晚期的乳腺癌患者,新辅助化疗也可以通过降低其分期达到保乳手术的目的。此外,新辅助化疗在保留乳腺、乳腺再建和淋巴结清扫手术中也有很大的益处。
阿霉素为乳腺癌化疗的常用药。阿霉素对肿瘤细胞的作用有两个主要机制:①嵌入DNA,破坏拓扑异构酶II介导的DNA修复过程;②破坏自由基的生成,以及它对细胞膜,DNA,蛋白质的损伤。然而一些体外研究表明,有些肿瘤细胞对阿霉素诱导凋亡作用具有很强的耐受性。阿霉素耐药重要机制主要包括:①ABCB1(P-gp)和ABCC1(MRP1)和其他转运蛋白(ABCC3、ABCG2、ABCC2)可以将多种抗癌药物排出细胞外。②TOP2A(DNA topoisomeraseII alpha)的扩增,这已被证明能够影响治疗的响应率。TOP2A扩增与邻近基因HER-2(ERBB2)有着错综复杂的关系。而HER-2作为乳腺癌治疗,特别是曲妥珠单抗治疗的一个标志。HER-2基因的扩增也会影响阿霉素治疗的响应率。③细胞膜脂质的生物物理性质及与药物的相互作用。研究指出,在阿霉素敏感细胞MCF-7和耐药细胞MCF-7/ADM模型中,耐药细胞膜脂质表现出明显不同的组成,更加凝聚的结构,流体膜少以及与与药物的强疏水作用等特点。耐药细胞的细胞膜刚性性质抑制了药物吸收,从而影响了化疗疗效。④microRNA影响阿霉素耐药:有研究发现,microRNA-451能调节MDR1的表达,在MCF-7/ADM耐药细胞中转染micro RNA-451有助于提高细胞对阿霉素的敏感性。⑤PI3K/AKT信号通路的激活。体外研究表明,PI3K/AKT的激活与阿霉素的化疗敏感性相关联。在乳腺癌治疗中结合常用的疗法,PI3K抑制剂LY294002增强了阿霉素、曲妥珠单抗、他莫昔芬引起的细胞凋亡。LY294002对化疗诱导的细胞凋亡的影响是通过AKT抑制导致的,这些研究表明,内源性AKT活性能促进乳腺癌患者的细胞存活和化疗耐药。
研究表明,阿霉素耐受处理的乳腺癌细胞对阿霉素有明显的耐受性,但具体的机理目前并不清楚。为了增加阿霉素对乳腺癌肿瘤细胞的杀伤作用,有必要寻找新的策略以逆转阿霉素耐受的乳腺癌细胞株对阿霉素的耐受性,从而达到阿霉素更有效治疗乳腺癌的目的。
川楝素(Toosendanin,TSN)是从传统中药川楝树的苦楝皮和川楝子中提取分离的一种四环三萜类化合物。作为天然中药药物成分,川楝素在驱虫、治疗肉毒毒素中毒、抗肿瘤、抑菌等多方面均显示出良好的生物活性和药理活性,极具开发研究价值。但是到目前为止,将川楝素用于肿瘤化疗增敏剂治疗乳腺癌并没有相关报道。
三、发明内容
阿霉素是目前临床上广泛使用的一种化疗药,是一种极具前景的抗癌药物。然而目前研究发现,许多恶性肿瘤细胞对阿霉素具有耐药性,使阿霉素在临床应用中遭遇瓶颈。川楝素在驱虫、治疗肉毒毒素中毒、抗肿瘤、抑菌等多方面均显示出良好的生物活性和药理活性。本发明中,我们首次发现川楝素能显著增强阿霉素耐药的乳腺癌肿瘤细胞株MCF-7/ADM对阿霉素诱导凋亡的敏感性。在其他的乳腺癌细胞株中,川楝素也能明显增强人源三阴性乳腺癌细胞MDA-MB-231,468细胞系以及鼠源乳腺癌细胞系4T1对阿霉素诱导凋亡的敏感性。免疫荧光法显示川楝素明显促进阿霉素在耐药细胞MCF-7/ADM中的积累,尤其是在细胞核中。进一步研究发现,川楝素能明显降低P-gp蛋白表达水平。动物实验也表明川楝素和阿霉素联用对肿瘤有很好的抑制作用。这些结果表明川楝素联用阿霉素,能有效增强阿霉素对耐受性乳腺癌肿瘤的治疗效果。
四、附图说明
图1 不同浓度的阿霉素对MCF-7和MCF-7/ADM凋亡作用
图2 川楝素增强MCF-7/ADM细胞对阿霉素的敏感性
图3 川楝素与阿霉素联用时MCF-7/ADM细胞中PARP的剪切水平
图4 川楝素和阿霉素对MDA-MB-231,MDA-MB-468,4T1凋亡作用
图5 免疫荧光检测川楝素对MCF-7/ADM细胞中阿霉素亚细胞分布的影响
图6 RT-PCR检测耐药相关基因在MCF-7/WT,MCF-7/ADM中的表达
图7 Western blotting检测P-gp在MCF-7,MCF-7/ADM中表达
图8 RT-PCR检测川楝素对耐药乳腺癌细胞中耐药相关基因表达的影响
图9 Western blotting检测川楝素处理时乳腺癌耐药细胞MCF-7/ADM中P-gp的蛋白表达
图10 Western blotting检测不同浓度川楝素处理对乳腺癌耐药细胞MCF-7/ADM中P-gp的蛋白表达的影响
图11 小鼠肿瘤组织
图12 小鼠肿瘤组织生长曲线
图13 小鼠肝脏和肾脏的HE染色
五、具体实施方式
5.1 实验材料、试剂
细胞:人源乳腺癌细胞株MCF-7和人源阿霉素耐药乳腺癌细胞株MCF-7/ADM购自中国医学科学院肿瘤医院肿瘤研究所,人源乳腺癌细胞株MDA-MB-231、
MDA-MB-468、鼠源乳腺癌细胞株4T1均来源于美国ATCC细胞库,以上细胞均用含10%的FBS、1%的链霉素青霉素双抗RPMI1640培养基培养。
分子试剂:川楝素Toosendanin(TSN,purity≥98%)(National Institutes forFood and Drug Control)、阿霉素(ADM)(生工生物工程(上海)股份有限公司)、Annexin-V(Invitrogen)、碘化丙碇(PI)(Invitrogen)、荧光染料Hoechst(碧云天)、tubulinantibody(Santa Cruz Biotechnology)、P-gp antibody(Abcam)、Goat Anti-Mouse IgG(santa cruz,SC-2005)、Goat Anti-rabbit IgG(santacruz,SC-2004)
5.2 PCR引物
用于半定量检测P-gp mRNA表达水平的引物为
RT P-gp F:5’-CTGGTTTGATGTGCACGATGTTGG-3’;
RT P-gp R:5’-TGCCAAGACCTCTTCAGCTACTG-3’
MRP1引物为
RT MRP1 F:5’-TCAGATGACACCTCTCAACAAAACC-3’;
RT MRP1 R:5’-GCTAGGGCACACACTAGGGCT-3’
BCRP引物为
RT BCRP F:5’-TCATGTTAGGATTGAAGCCAAAGGC-3’;
RT BCRP R:5’-TGTGAGATTGACCAACAGACCTGA-3’
内参引物为
RT GAPDH F:5’-CACCATCTTCCAGGAGCGAG-3’;
RT GAPDH R:5’-GCAGGAGGCATTGCTGAT-3’
5.3 实验方法:
5.3.1. 流式细胞仪检测细胞凋亡
5.3.1.1样品制备:
1)培养待测细胞至汇合度约60%左右,给药处理细胞;
2)处理细胞达到指定时间,胰酶消化法收集贴壁细胞。将收集的细胞重悬于预冷的PBS缓冲液中洗涤1-2次,2500rpm离心5min,弃上清;
3)用缓冲液(1×binding buffer)洗涤细胞1-2次,2500rpm离心5min,弃上清;
4)配制Annexin V染液,用1×binding buffer配制Annexin V-EGFP染液,使其终浓度为1ug/ml;用400μL的染液重悬细胞,移至冰上避光孵育15-30min;
5)向每管样品中加入适量PI(碘化丙碇),使其终浓度为1ug/ml,冰上避光孵育5min。
6)将细胞转移至流式管,用流式细胞仪上机分析。
5.3.1.2流式细胞仪分析
1)打开流式细胞仪和计算机。打开流式细胞仪检测软件,流式细胞仪的激发光波长为488nm的氩离子镭射,Annexin V-EGFP发出的绿色荧光可以用FLI通道检测,PI(碘化丙碇)受激发所发出的红色荧光用FL3通道检测;
2)建立FSC-SSC散点图,并用自由门工具圈出大部分细胞群体;
3)以门中的细胞群建立散点图,横纵坐标分别为FL1和FL3通道。在散点图上设置十字门,将得到的散点图分为4个象限。正常情况下的细胞在流式细胞仪分析时可以分成三个亚群:A:正常活细胞(AnnexinV、PI亲和度低,位于左下象限);B:早期凋亡细胞(AnnexinV亲和度高、PI亲和度低,位于右下象限);C:中晚期凋亡细胞或坏死细胞(AnnexinV、PI亲和度高,位于右上象限);调节相关参数,使对照组细胞的绝大部分细胞处于左下象限区域;
4)依次检测实验组中不同处理的细胞。
5.3.1.3凋亡数据处理和分析
凋亡实验中每个处理的样品都设置三个以上的重复,最终数据均mean SEM表示。数据进行差异统计时采用student’s t test检验,当p≥0.05时认为没有显著性差异(notsignificant,ns);p<0.05时差异显著(*),p<0.01时差异极显著(**),p<0.001时差异极显著(***)。
5.3.2 细胞免疫荧光
细胞铺于含细胞爬片的12孔板中,密度大约40%-50%,培养、加药处理。①弃去培液,预冷PBS洗3次。4%多聚甲醛固定细胞,室温孵育30min。②用预冷PBS洗3次,每次5min。加入0.5%TritonX-100室温孵育10min。③用预冷PBS洗3次,每次5min。④滴加Hoechst(1∶10000,用PBS稀释),染色10min,用预冷PBS洗3次,每次5min。⑤封片:取出12孔板中的细胞爬片,滴加适量甘油于载玻片上,并将爬片倒扣于载玻片上,荧光显微镜下观察。
5.3.3. 半定量PCR(RT-PCR)
5.3.3.1 总RNA的提取
1)准备工作:所有实验用品需为RNase-free,用DEPC处理并灭菌。
2)收细胞:用预冷的PBS洗涤细胞一遍,加适量的TRIZOL agent充分吹打均匀,转移至EP管中,室温静置5min。
3)加入TRIZOL体积1/5的氯仿,剧烈震荡15s充分混匀,室温静置5min。
4)12000rpm,4℃离心15min。离心完毕,EP管中的液体分为三层。
5)小心将最上层澄清相转移至另一EP管中,加入等体积的异丙醇,颠倒混匀,室温静置10min。
6)12000rpm,4℃离心15min。离心完毕,可见RNA沉淀,弃上清。
7)缓缓沿EP管壁加入适量的75%乙醇(DEPC水配制),轻轻颠倒洗涤沉淀。
8)12000rpm,4℃离心5min,弃去乙醇,晾干RNA沉淀。
9)加入30μL DEPC水溶解沉淀,可于60℃水浴中助溶。
10)紫外分光光度计测定RNA浓度,并加DEPC水调整RNA浓度至所有样品浓度一致。
5.3.3.2 逆转录PCR合成cDNA
采用10μL的逆转录体系:
配好后充分混匀,离心,进行逆转录反应:37℃,30min;98℃,5min。将扩增好的cDNA按适当比例稀释,-80℃长期保存。
5.3.3.3 半定量PCR(RT-PCR)
采取20μL的配制扩增体系:
充分混匀,离心,并进行PCR反应。
PCR条件如下:
PCR产物加入适量的DNA上样缓冲液,1%琼脂糖凝胶电泳分析,EB显色。
5.3.4 Western Blotting
1)SDS-PAGE胶的配制:根据《分子克隆》中的配方配制8-15%蛋白分离胶以及5%的蛋白浓缩胶;
2)细胞样品经过不同药物处理后用RIPA裂解液裂解,用BCA比色法测定的样品中的蛋白浓度,并将所有样品的蛋白浓度调整一致,加入适量的蛋白上样缓冲液,充分混匀,煮沸5min使蛋白变性,-80℃长期保存;
3)根据《分子克隆》中的配方配置电泳液和转膜液,转膜液置于4℃展示柜预冷;
4)电泳:各组蛋白样品上样,上样量需保持一致(10-40μg)。电泳的前30min采用80V恒压使得蛋白样品在浓缩胶中进行压缩。随后提高电压至120V进行分离;
5)转膜:使用湿转法将SDS-PAGE胶中的蛋白转印至PVDF膜上。根据目的蛋白的分子量大小选择适合孔径的PVDF膜,300mA恒流电转1.5-2h;根据蛋白Marker的转印效果判断转印效率;
6)封闭:转印完毕后,将PVDF膜放入现配的含有5%脱脂奶粉的PBST中,室温封闭30min左右;
7)孵育一抗:PVDF膜封闭结束后,根据目的蛋白对应抗体说明书使用5%BSA溶液稀释一抗至对应的最佳工作浓度,将PVDF膜包被于稀释好的一抗溶液中,室温孵育2h或4℃过夜;
8)孵育二抗:回收一抗,将PVDF膜取出置于适量PBST中,在摇床上以适当转速洗涤3次,每次5min;同时,将由HRP标记的对应一抗种属的二抗按照说明书稀释于5%的BSA溶液中;洗涤完毕,将PVDF膜包被于二抗中,室温孵育1h;
9)显影:二抗孵育完毕,以PBST将PVDF膜洗涤3次,每次5min。同时,按照说明书配制ECL化学发光液。洗涤完毕后将PVDF膜取出,吸掉膜上的PBST并置于化学发光仪的暗箱中,将适量化学发光液滴于膜上,关闭暗箱箱门,在软件上设置适当的曝光时间进行蛋白检测。
5.3.5 4T1肿瘤细胞移植瘤模型的建立及给药处理
1)种4T1细胞于20cm的细胞培养皿中,待其状态良好,密度约为80%左右时消化收集细胞,用预冷的PBS洗涤两次,用适量PBS重悬并计数。
2)接种5×106个/0.2mL的4T1细胞于BALB/C雌性小鼠的右侧腹部的脂肪垫中。
3)给药处理小鼠,给药方式为尾静脉注射,每两天注射一次。
4)每两天测量一次肿瘤大小,肿瘤体积计算公式为:1/2×最长径×最短径×最短径。
5)15天后颈椎脱臼法处死小鼠,取肝、肾和肿瘤组织,用福尔马林溶液固定,送谷歌生物制作石蜡切片。
5.3.6 石蜡切片的H&E染色
1)石蜡切片的脱腊和水化:二甲苯I 15min,二甲苯II 15min,无水乙醇5min,95%乙醇I 5min,95%乙醇II 5min,80%乙醇5min,DDW 10min。
2)苏木素染色10-15min,流水冲洗1-3s。
3)分化液分化1-3s。
4)促蓝液返蓝30s,稍水洗。
5)伊红染液染色5min,稍水洗。
6)脱水:无水乙醇 5min,二甲苯II 15min,二甲苯I 15min。
7)中性树胶封片,显微镜观察。
5.4 实验结果:
实施例一 川楝素增强乳腺癌细胞MCF-7/ADM对化疗药物阿霉素的敏感性
取生长状态良好的对数生长期人乳腺癌细胞MCF-7,阿霉素耐药的乳腺癌细胞MCF-7/ADM种植于24孔板,待细胞贴壁生长良好后用含不同浓度阿霉素的培养液处理36h,收集细胞并用流式细胞术测定细胞凋亡情况。结果显示,耐药细胞株MCF-7/ADM对阿霉素表现出很强的耐药性(图1)。然后我们对MCF-7/ADM细胞进行单独加药和联合加药处理。我们将MCF-7/ADM细胞分4组:对照组,阿霉素(10μg/ml)组,川楝素(200nM)组,阿霉素(10μg/ml)+川楝素(200nM)组,加药处理36h后流式细胞术检测细胞凋亡。结果表明,用阿霉素和川楝素单独处理后,MCF-7/ADM表现出显著的耐药性,细胞没有产生明显的凋亡(凋亡率5%左右)。然而,当阿霉素与川楝素联合使用时,MCF-7/ADM细胞表现出明显的凋亡(凋亡率达60%),以上结果揭示出川楝素能显著增强乳腺癌耐药细胞对阿霉素敏感性(图2)。
实施例二 川楝素与阿霉素联用细胞PARP剪切水平增加
细胞内PARP蛋白发生剪切通常视为细胞凋亡的重要标志。在我们的研究中,利用Westernblotting检测了川楝素与阿霉素联合用药诱导阿霉素耐药MCF-7/ADM细胞中PARP的剪切水平。结果表明,不同浓度川楝素联用阿霉素(20μg/ml)处理后,随着川楝素的浓度增加,细胞中的PARP剪切水平增强。结果表明,川楝素与阿霉素联合用药时能显著增强阿霉素诱导的耐药细胞MCF-7/ADM细胞发生凋亡,并且这种促凋亡作用与川楝素的浓度呈正相关(图3)。
实施例三 川楝素能增加不同类型的乳腺癌细胞对阿霉素的药物敏感性
为了研究川楝素对乳腺癌细胞阿霉素耐药性的逆转作用是否具有普遍性,我们进一步验证了川楝素与阿霉素联用对另三种乳腺癌细胞(人源三阴性乳腺癌细胞MDA-MB-231,MDA-MB-468细胞系以及鼠源乳腺癌细胞系4T1)阿霉素耐药的作用。结果显示,在MDA-MB-231以及468细胞中,川楝素与阿霉素联用与阿霉素单用相比,能明显增加细胞的凋亡(约2-3倍)。而在4T1细胞中,川楝素与阿霉素联用时,即使川楝素在较低浓度(25nM)时,其诱导的细胞凋亡率比单用阿霉素增加4-5倍,显示出其良好的增敏效应(图4)。
实施例四 川楝素促进阿霉素在乳腺癌耐药细胞中的积累
乳腺癌细胞MCF-7,MCF-7/ADM制备细胞爬片,待细胞贴壁生长良好后,用无毒剂量的川楝素提前处理12h,然后加10μg/ml的阿霉素处理6h后进行免疫荧光染色。结果表明,MCF-7细胞核内阿霉素的荧光强度明显高于MCF-7/ADM细胞,说明MCF-7/ADM细胞内的阿霉素积累量明显低于MCF-7细胞。而川楝素处理组的MCF-7/ADM耐药细胞株细胞核中阿霉素荧光强度明显高于溶剂对照组细胞胞核中阿霉素的荧光强度,表明川楝素可以明显增加阿霉素在MCF-7/ADM中的聚集和积累(图5)。
实施例五 川楝素下调乳腺癌耐药细胞中P-gp的蛋白表达
我们进一步探究了川楝素逆转乳腺癌细胞对阿霉素的耐受性的分子机制。取对数生长期细胞MCF-7,MCF-7/ADM,RT-PCR或Western blotting检测细胞中耐药相关基因以及蛋白的表达情况。结果显示,与MCF-7相比,MCF-7/ADM细胞中P-gp,MRP1,BCRP基因的mRNA表达水平显著升高(图6)。Western blotting结果显示,在MCF-7/ADM细胞中耐药蛋白P-gp高表达,而在MCF-7细胞中基本不表达(图7)。
我们进一步研究了川楝素对MCF-7/ADM细胞中P-gp表达量是否有影响。取对数生长期的MCF-7/ADM细胞铺板,用不同浓度川楝素处理细胞36h,RT-PCR检测细胞中P-gp的mRNA水平。结果显示,在无毒剂量的川楝素浓度下,P-gp的mRNA表达量基本没有发生变化(图8)。然而western blotting结果显示,川楝素能明显下调MCF-7/ADM细胞中P-gp的蛋白表达(图9)。通过进一步实验我们发现,川楝素下调P-gp的蛋白表达存在一定的浓度依赖性,随着川楝素的浓度增大,抑制效果也在不断增加(图10)。
实施例六 川楝素和阿霉素联用抑制小鼠体内肿瘤的生长
为了验证川楝素和阿霉素联用在体内的作用,我们把4T1细胞注射到BALB/C雌性小鼠的右侧腹部的脂肪垫中。小鼠随机分为4组,每组6只。每隔一天分别通过尾静脉注射PBS,阿霉素(4mg/kg),川楝素(0.69mg/kg),阿霉素(4mg/kg)+川楝素(0.69mg/kg),每两天测量小鼠的肿瘤大小。15天后颈椎脱臼法处死小鼠。结果表明,阿霉素和川楝素单独注射,对小鼠肿瘤生长有微弱的抑制作用。而阿霉素和川楝素联合注射对小鼠肿瘤具有显著的抑制作用(图11,12)。而HE结果表明,我们使用的川楝素剂量对小鼠的肝脏和肾脏都没有毒害作用(图13)。实验表明,川楝素和阿霉素联用对动物体内的乳腺癌细胞具有很好的抑制作用。
Claims (3)
1.川楝素在制备抗乳腺癌化疗药物增敏剂中的应用。
2.根据权利要求1所述的应用,所述川楝素的结构为:
3.根据权利要求1所述的应用,其特征在于:用川楝素制备的药物以注射剂或输液剂给药。
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CN115137734A (zh) * | 2022-08-04 | 2022-10-04 | 苏州大学 | 一种用于治疗耐药性癌症的药物组合物 |
CN115177745A (zh) * | 2022-08-11 | 2022-10-14 | 福建医科大学附属口腔医院 | 一种川楝素及其纳米材料复合物的制备方法和应用 |
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CN114940696A (zh) * | 2022-05-25 | 2022-08-26 | 四川大学华西医院 | 一种川楝素衍生物及其在乳腺癌治疗中的应用 |
CN114940696B (zh) * | 2022-05-25 | 2023-04-28 | 四川大学华西医院 | 一种川楝素衍生物及其在乳腺癌治疗中的应用 |
CN115137734A (zh) * | 2022-08-04 | 2022-10-04 | 苏州大学 | 一种用于治疗耐药性癌症的药物组合物 |
CN115177745A (zh) * | 2022-08-11 | 2022-10-14 | 福建医科大学附属口腔医院 | 一种川楝素及其纳米材料复合物的制备方法和应用 |
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