Antiproliferative effects of cinobufacini on human hepatocellular carcinoma HepG2 cells detected by atomic force microscopy

AIM:To investigate the antiproliferative activity of cinobufacini on human hepatocellular carcinoma HepG2 cells and the possible mechanism of its action.METHODS:HepG2 cells were treated with different concentrations of cinobufacini.Cell viability was measured by methylthiazolyl tetrazolium(MTT) assay.Cell cycledistribution was analyzed by flow cytometry(FCM).Cytoskeletal and nuclear alterations were observed by fluorescein isothiocyanate-phalloidin and DAPI staining under a laser scanning confocal microscope.Changes in morphology and ultrastructure of cells were detected by atomic force microscopy(AFM) at the nanoscale level.RESULTS:MTT assay indicated that cinobufacini significantly inhibited the viability of HepG2 cells in a dosedependent manner.With the concentration of cinobufacini increasing from 0 to 0.10 mg/m L,the cell viability decreased from 74.9% ± 2.7% to 49.41% ± 2.2% and 39.24% ± 2.1%(P < 0.05).FCM analysis demonstrated cell cycle arrest at S phase induced by cinobufacini.The immunofluorescence studies of cytoskeletal and nuclear morphology showed that after cinobufacini treatment,the regular reorganization of actin filaments in HepG2 cells become chaotic,while the nuclei were not damaged seriously.Additionally,high-resolution AFM imaging revealed that cell morphology and ultrastructure changed a lot after treatment with cinobufacini.It appeared as significant shrinkage and deep pores in the cell membrane,with larger particles and a rougher cell surface.CONCLUSION:Cinobufacini inhibits the viability of HepG2 cells via cytoskeletal destruction and cell membrane toxicity.

Impact of Cinobufacini injection on proliferation and cell cycle of human hepatoma HepG-2 cells

Objective： The aim of our study was to investigate the effect of Cinobufacini injection on the proliferation and cell cycle of human hepatoma HepG-2 cells. Methods： Cell proliferation was assessed by MTT assay, cell cycle distribution was detected by the flow cytometry （FCM）. The expression of Cyclin A, CDK2 mRNA levels were examined by RT-PCR. Quantitative colorimetric assay was used to analyze Cyclin NCDK2 activity in HepG-2 cells. Results： Cinobufacini injection significantly inhibited HepG-2 cells proliferation in dose- and time-dependent ways; FCM analysis showed Cinobufacini injection induced cell cycle arrest at S phase; RT-PCR assay showed Cinobufacini injection down-regulated Cyclin A, CDK2 expression at mRNA levels; Quantitative colorimetric assay showed Cinobufacini injection deceased Cyclin NCDK2 activity in HepG-2 cells. Conclusion： Cinobufacini injection can inhibit human hepatoma HepG-2 cells growth, induce cell apoptosis and induce cell cycle arrest at S phase, the mechanism of which might be partly related to the down-regulation of Cyclin A, CDK2 mRNA expression and inhibition of Cyclin A/CDK2 activity.

The impact of Cinobufacini injection on proliferation and apoptosis of human hepatoma HepG-2 cells

Objective： The aim of our study was to investigate the effect of Cinobufacini injection on the proliferat（on and apoptosis of human hepatocarcinoma HepG-2 cells. Methods： Cells proliferation was assessed by MTT assay, cells morphologic was observed by the inverted microscopy, Annexin V/PI stain was used to detect the apoptosis and necrosis of the tumor ceils. The expression of TOPOI mRNA and TOPO Ⅱ mRNAwere examined by RT-PCR. Results： Cinobufacini injection significantly inhibited HepG-2 cells proliferation in dose- and time-dependent ways. After Cinobufacini injection intervention, HepG-2 cells showed typical morphological changes： cells changed from polygon into round, chromatin looseness and karyolysis were observed. The percentages of apoptosis were 88.49%, 76.02%, 61.73% corresponding to the 48 h interference of 0.42 μg/mL, 0.21 μg/mL, 0.105 μg/mL Cinobufacini injection, perspectively. RT-PCR assay showed that Cinobufacini injection down-regulated TOPOI and TOPO Ⅱ expression at mRNA level. Conclusion： Cinobufacini can inhibit human hepatocarcinoma HepG-2 cells growth and induce tumor cells apoptosis, the mechanism of which might partly relate to the down-regulation of TOPOI mRNA and TOPO Ⅰ mRNA induced by Cinobufacini injection.

Chemical Composition, Pharmacological Effects and Clinical Applications of Cinobufacini

Chinese medicine cinobufacini is an extract from the dried skin of Bufo bufo gargarizans Cantor,with active ingredients of bufadienolides and indole alkaloids.With further research and clinical applications,it is found that cinobufacini alone or in combination with other therapeutic methods can play an anti-tumor role by controlling proliferation of tumor cells,promoting apoptosis,inhibiting formation of tumor neovascularization,reversing multidrug resistance,and regulating immune response;it also has the functions of relieving cancer pain and regulating immune function.In this paper,the chemical composition,pharmacological effects,clinical applications,and adverse reactions of cinobufacini are summarized.However,the extraction of monomer components of cinobufacini,the relationship between different mechanisms,and the causes of adverse reactions need to be further studied.Also,high-quality clinical studies should be conducted.

Cinobufacini injection for moderate and advanced primary liver cancer: A systematic review and meta-analysis

Cinobufacini, a compound extracted from bufo toad, is a traditional Chinese medicine(TCM) that has been shown to have efficacy in cancer treatment.The cinobufacini injection is widely used in patients with moderate and advanced liver cancer in China.In the present study, we aimed to determine the effects of cinobufacini injection for this specific category of patients.A search for randomized control trials(RCTs) using cinobufacini was conducted in the PubMed, EMBASE, CENTRAL, and the other four major Chinese databases.The systematic review was performed according to the recommendations of the Cochrane collaboration, and the RevMan 5.3.software was used for statistical analysis.A random-effects model was used to perform the data.Risk ratios(RR) with corresponding 95% confidence interval were calculated according to Cochrane handbook.A total of 11 RCTs consisting of 728 patients were included.All of the trials demonstrated significantly improved total response rates, total response rates of Karnofsky Performance Score(KPS), 1-to 2-year survival rates, and quality of life in the intervention groups injected with cinobufacini.There was no statistically significant difference between the groups(cinobufacini versus no cinobufacini; cinobufacini plus transcatheter arterial chemoembolization(TACE) versus TACE only) in terms of the 6-month survival rate, clinical benefit rate and clinical benefit rate of KPS.This systematic review demonstrated the beneficial effects of cinobufacini injection in terms of total response rate and the 1-to 2-year survival rate in patients with moderate and advanced liver cancer.The efficacy for the cinobufacini injection group might be better than that of the control group for treatment of moderate and advanced liver cancer.Given that the majority of the trials were of low quality, more high-quality prospective RCTs with strict design in accordance with CONSORT for TCM are needed.

华蟾素通过调控MYH9/USP7/c-MYC通路抑制急性髓系白血病细胞免疫逃逸

【目的】探讨华蟾素调控肌球蛋白重链9(MYH9)/泛素特异性蛋白酶7(USP7)/骨髓细胞瘤病毒癌基因(c-MYC)通路对急性髓系白血病(AML)细胞免疫逃逸的影响。【方法】(1)体内实验:建立裸鼠异种移植瘤模型,评估华蟾素对AML细胞在体内生长和免疫逃逸的影响。(2)体外实验:使用不同浓度的华蟾素处理人AML细胞株HL-60,细胞计数试剂盒8(CCK-8)法检测细胞活力,Transwell实验检测细胞侵袭能力。将HL-60细胞与活化的CD8^(+)T细胞共培养,流式细胞术检测CD8^(+)T细胞表面标志物CD25的表达,酶联免疫吸附分析(ELISA)检测共培养上清液中细胞因子[白细胞介素2(IL-2)和干扰素(IFN-γ)]的水平,CytoTox96非放射性细胞毒性分析评估CD8^(+)T细胞对HL-60细胞的毒性。Western Blot法检测MYH9、USP7和c-MYC的蛋白表达,免疫共沉淀(Co-IP)法检测MYH9、USP7和泛素化之间的相互作用。转染MYH9过表质粒,验证华蟾素在AML中的作用机制。【结果】华蟾素抑制裸鼠移植瘤生长,增强CD8^(+)T细胞抗肿瘤的能力。华蟾素浓度依赖性地抑制HL-60细胞活力、侵袭。华蟾素处理后上调CD8^(+)T细胞表面标志物CD25的表达,同时还上调IL-2和IFN-γ水平。华蟾素增强CD8^(+)T细胞对HL-60细胞的毒性。华蟾素抑制HL-60细胞MYH9、USP7和c-MYC的蛋白表达,MYH9通过募集USP7促进c-MYC去泛素化,华蟾素抑制MYH9介导的c-MYC去泛素化。【结论】华蟾素可通过抑制MYH9的表达进而减少去泛素化酶USP7对c-MYC的募集,促进c-MYC泛素化降解,从而抑制AML细胞免疫逃逸。

华蟾素通过调控M2型巨噬细胞极化抑制结直肠癌转移

目的探讨华蟾素通过调控M2型巨噬细胞极化抑制结直肠癌转移的作用。方法将THP-1诱导成M0型巨噬细胞,收集HCT116细胞的条件培养基,刺激M0型巨噬细胞,通过流式细胞术、RT-qPCR、ELISA实验观察M2型巨噬细胞极化状态;收集M0型巨噬细胞和HCT116-Mφ细胞的条件培养基,刺激HCT116细胞,通过划痕实验和Transwell实验观察迁移和侵袭能力;用CCK-8实验检测华蟾素对HCT116细胞活力的影响;收集HCT116和HCT116+华蟾素的条件培养基,刺激M0型巨噬细胞,通过流式细胞术、RT-qPCR、ELISA实验观察M2型巨噬细胞极化状态;收集HCT116-Mφ细胞和(HCT116+华蟾素)-Mφ细胞的条件培养基,刺激HCT116细胞,通过划痕实验和Transwell实验观察迁移和侵袭能力的改变。结果M0型巨噬细胞在HCT116细胞的条件培养基刺激后,形态变成梭形的细胞,CD11b^(+)CD206^(+)细胞比例增高,M2型巨噬细胞标志物白细胞介素-10(IL-10)及转化生长因子-β(TGF-β)表达升高;HCT116细胞在HCT116-Mφ细胞的条件培养基刺激后,细胞迁移和侵袭能力明显增强;加入华蟾素之后,不仅M2型巨噬细胞极化比例降低,M2型巨噬细胞介导的促转移效应也受到抑制。结论HCT116细胞可以诱导M2型巨噬细胞极化,而华蟾素可通过抑制M2型巨噬细胞极化,进而抑制M2型巨噬细胞介导的肿瘤转移。

Preventive Effects of Jiedu Granules(解毒颗粒) Combined with Cinobufacini Injection(华蟾素注射液) versus Transcatheter Arterial Chemoembolization in Post-Surgical Patients with Hepatocellular Carcinoma:A Case-Control Trial

Objective： To investigate the therapeutic effects of Jiedu Granules （解毒颗粒）, a Chinese medicine （CM） compound, plus Cinobufacini Injection （华蟾素注射液）, which was extracted from skin of Bufo bufo gargarizans Cantor, to prevent the recurrence of hepatocellular carcinoma （HCC） after surgical resection. Methods： In this case-control trial, a total of 120 patients who stayed in Changhai Hospital were enrolled from December 2001 to December 2006. Sixty patients were treated with Jiedu Granules plus Cinobufacini Injection to prevent tumor recurrence after operation （CM group） and 60 patients were treated with transcatheter arterial chemoembolization （TACE） after operation （TACE group）. Progression-free survival （PFS） and overall survival （OS） rates were determined to evaluate the therapeutic effects of post-operative management of patients with HCC. Results： PFS in the CM group was 18.07 months [95% confidence interval （CI）： 12.49-23.65] and the 1-, 2-, 3-, 4- and 5-year PFS rates were 61%, 39%, 26%, 22% and 12%, respectively. PFS in the TACE group was 8.03 months （95% CI： 6.63-9.44） and the 1-, 2-, 3-, 4- and 5-year PFS rates were 34%, 11%, 7%, 2% and 0%, respectively. There was significant difference in survival rate between the two groups （P〈0.01）. The mean survival time （MST） of patients in the CM group was 49.53 months versus 39.90 months of the TACE group. The 1-, 2-, 3-, 4- and 5-year survival rates were 90%, 82%, 80%, 70% and 63%, respectively, in the CM group, and 79%, 70%, 60%, 60% and 36%, respectively, in the TACE group. There was significant difference in survival time between the two groups （P=0.045）. Conclusions： Jiedu Granules plus Cinobufacini Injection, a combination that is commonly used for post-operation management of HCC, can postpone tumor recurrence and metastasis, prolong the survival time and increase the survival rate of post-surgical patients with HCC. However, these findings need to be confirmed in a prospective, randomized controlled trial.

Apoptosis of T-47D Cells Induced by Cinobufacini via a Caspase-3-dependent Manner

To investigate the mechanism of the anti-tumor activity of cinobufacini on the breast cancer cell line T-47D,the inhibitory effect of cinobufacini on the proliferation of T-47D was detected via MTT assay and the morphological changes of T-47D and HBL-100 cells caused by cinobufacini were observed with an inverted microscope.Cell apoptosis and cell cycle stages were detected by flow cytometry analysis.The effects of cinobufacini on the expression of active-form and pro-form of caspase-3 were assessed by Western blot analysis.Cinobufacini dramatically inhibited T-47D proliferation in a dose-and time-dependent manner.We found that more than 20％ of T-47D cells were killed after treatment with 20 mg/mL cinobufacini for 24 h in vitro.After 6 d of treatment with 20 mg/mL cinobufacini,the cell survival rate decreased by more than 40％.Flow cytometric analysis demonstrated that cinobufacini induced significant apoptosis and changes of the cell cycle distribution of T-47D cells.We used breast cell line HBL-100 as the control,the above experiments except cell cycle analysis showed that cinobufacini more obviously induced the apoptosis of T-47D cells than that of HBL-100 cells.Western blot analysis confirmed the protein expression of active caspase-3 increased with increasing the dose of cinobufacini.These results indicate that cinobufacini induces the apoptosis of T-47D cells via the up-regulation of caspase-3.

基于网络药理学探讨华蟾素对肺癌的作用机制

目的:通过网络药理学方法系统地探讨华蟾素抗肺癌的作用机制。方法:利用文献资料和比较毒理学基因组数据库(CTD)获取华蟾素的活性组分,再用SwissADME数据库对其有效成分进行分析后,利用UniProt数据库对其基因命名作规范化处理。从GeneCards数据库以及OMIM数据库获取肺癌基因;利用韦恩图得到华蟾素抗肺癌的潜在治疗靶点;通过STRING数据库来获得潜在治疗靶点的蛋白质相互作用,后用Cytoscape软件创建蛋白质-蛋白质相互作用(PPI)网络并确定关键作用靶点;使用Cytoscape软件创建药物-成分-靶点-疾病的网络,并筛出关键活性成分;用Metascape数据库对潜在治疗靶点作GO功能富集和KEGG通路富集,之后用Cytoscape软件创建药物-靶点-通路的网络。结果:华蟾素含有49种活性化合物,活性成分靶基因195个,肺癌靶基因共1 197个,华蟾素抗肺癌潜在治疗靶点56个。GO功能富集891个条目,KEGG通路富集分析共136个条目。结论:华蟾素通过多种活性成分、多靶点、多通路相互作用发挥抗肺癌作用。

华蟾素片联合DP化疗方案治疗中晚期非小细胞肺癌对患者免疫功能、血清肿瘤标志物水平影响

目的:探讨华蟾素片联合多西他赛+顺铂(DP)化疗方案治疗中晚期非小细胞肺癌(NSCLC)的效果。方法:随机抽选龙岩市第二医院2020年12月—2021年12月收治的中晚期NSCLC患者,共90例,依照随机数字表法分为参考组(45例)和研究组(45例)。参考组采用DP化疗方案治疗,研究组在DP化疗方案的基础上联合华蟾素片治疗。观察两组治疗前后免疫功能指标(CD3^(+)T细胞、CD4^(+)T细胞、CD8^(+)T细胞、CD4^(+)/CD8^(+))和肿瘤标志物[血管内皮生长因子(VEGF)、癌胚抗原(CEA)、神经元特异性烯醇化酶(NSE)]水平的变化情况。结果:治疗后,两组CD3^(+)T细胞、CD4^(+)T细胞和CD4^(+)/CD8^(+)均较治疗前升高,且研究组上述指标均明显高于参考组,两组CD8^(+)T细胞均较治疗前下降,研究组CD8^(+)T细胞明显低于参考组(P<0.05)。治疗后,两组VEGF、CEA和NSE均下降,研究组的VEGF、CEA和NS均低于参考组(P<0.05)。结论:对中晚期NSCLC患者采用华蟾素片联合DP化疗方案治疗,可显著提升患者免疫功能,降低血清肿瘤标志物表达,与单纯DP化疗相比更有优势。

华蟾素注射液中蟾毒内酯类成分含量检测

目的 :用高效液相色谱法分离测定华蟾素注射液中蟾毒灵、华蟾酥毒基、脂蟾毒配基等成分含量。方法 :醋酸乙酯分离提取华蟾素中脂溶性成分 ,利用高效液相色谱仪定量检测 ,以Econosphere C1 8柱为分析柱 ,乙腈 水 (50∶50 )为流动相 ,检测波长 2 99nm。结果 :该测定方法准确 ,分离效果好 ,灵敏度高。根据实验结果计算华蟾素注射液中各蟾毒内酯类成分含量分别为蟾毒灵 0 .333μg·mL- 1 ,华蟾酥毒基 0 .1 59μg·mL- 1 ,脂蟾毒配基 0 .1 1 0 μg·mL- 1 。结论 :华蟾素注射液中蟾毒灵浓度从理论上讲已达到有效作用浓度 ,可能是华蟾素注射液抗肿瘤有效成分之一。

华蟾素注射液治疗中晚期原发性肝癌临床疗效观察

目的 观察华蟾素注射液治疗中晚期原发性肝癌的临床疗效。方法 10 0例中晚期原发性肝癌患者随机分成华蟾素治疗组与一般治疗组 ,各 50例。分别观察两组患者治疗前后的生存质量、肿瘤大小、部分实验室指标的变化 ,并随访生存期。结果 在控制肿瘤方面 ,华蟾素治疗组的恶化率 (18% )低于一般治疗组 (3 2 % ) ;在生存质量方面 ,华蟾素治疗组的总有效率(80 % )高于一般治疗组 (72 % ) ,但两组比较无显著性差异 ;在生存期方面 ,华蟾素治疗组 >12个月的生存率 (3 0 % )显著高于一般治疗组 (18% ) ,两组比较有显著性差异 ;在肝功能方面 ,华蟾素治疗组的血清总胆红素、丙氨酸氨基转移酶等指标在治疗后有明显下降 ,而在一般治疗组则呈不同程度的升高 ;在甲胎蛋白 (AFP)方面 ,一般治疗组的AFP在治疗后较治疗前有明显上升 ,而华蟾素治疗组则略有上升 ,但与治疗前相比无显著性差异。结论 华蟾素注射液能在一定程度上抑制癌瘤的生长 ,并且具有保护肝功能、提高生存质量、延长生存期的作用 ,在治疗中晚期原发性肝癌方面 ,具有“攻邪不伤正”

三氧化二砷联合华蟾素抗裸鼠人肝癌移植瘤血管新生的作用

目的探讨三氧化二砷(arsenic trioxide,As2O3)联合华蟾素对裸鼠人肝癌移植瘤血管新生的作用及其相关机制。方法建立裸鼠肝癌原位移植瘤模型,随机分为4组,即生理盐水组(对照组)、As2O3组、华蟾素组和As2O3联合华蟾素组(联合组),每组8只,成瘤后腹腔内分别连续21天注射生理盐水、2.5mg/kg As2O3注射液、5mL/kg华蟾素注射液和2.5mg/kg As2O3注射液+5mL/kg华蟾素注射液。比较各组裸鼠一般状态、移植瘤大小、肿瘤微血管密度(microvessel density,MVD),采用免疫组化、光镜、透射电镜进行观察移植瘤血管内皮生长因子(vascular endothelial growth factor,VEGF)、表皮生长因子受体(epidermalgrowth factor receptor,EGFR)及移植瘤病理,并对裸鼠肝、肾组织病理学和血常规进行检测。结果 As2O3组、华蟾素组及联合组的平均瘤重(g,0.65±0.25、0.70±0.27、0.42±0.16)及平均瘤体积(cm3,0.44±0.14、0.46±0.19、0.26±0.11)均低于对照组平均瘤重(1.06±0.25)及平均瘤体积(0.67±0.17,P<0.05);两药相互作用系数(coefficient of drug interaction,CDI)值计算结果为:瘤重CDI(0.97)<1,体积CDI(0.86)<1,表明两药有协同抑制移植瘤生长的作用。As2O3和华蟾素均能抑制移植瘤VEGF、EG-FR的表达,并可降低肿瘤MVD,两药联合后上述作用均显著增强(P<0.05)。裸鼠原位移植瘤的光镜和电镜下观察结果显示As2O3、华蟾素及两药联合均可抑制原位移植瘤组织细胞生长。联合用药并未增加裸鼠肝、肾和造血系统的毒性。结论 As2O3联合华蟾素协同抑制裸鼠人肝癌移植瘤的血管新生,能有效抑制移植瘤生长;同时联合用药对于荷瘤裸鼠的肝、肾和造血系统未见明显毒性。

不同护理方法防治华蟾素所致静脉炎的效果观察

中晚期原发性肝癌常因无法手术或行肝动脉化疗栓塞而得不到有效治疗。目前临床采用中西医结合治疗中晚期原发性肝癌，取得了一定疗效。华蟾素注射液是常用的中药抗肿瘤制剂。临床研究表明，其不仅能抗癌祛邪，且能减毒扶正，从而起到抑制肿瘤生长、提高生存质量和延长生存期的作用，被广泛应用于中晚期原发性肝癌的治疗。但应用过程中，常因出现静脉滴注局部红、肿、疼痛等静脉炎症状，使患者不愿接受或不能坚持长期治疗，从而影响治疗效果。

AO/EB染色观察华蟾素诱导体外肿瘤细胞凋亡

目的:观察华蟾素诱导肿瘤细胞凋亡的变化规律,找出最佳诱导剂量及最佳诱导时间.方法:采用人肝癌SMMC-7721和人胃癌MGC-80-3细胞株,以AO/EB染色对华赡素不同浓度及作用时间下细胞凋亡的形态变化进行观察,并定量计算出凋亡率.结果:在荧光显微镜下可区分早期凋亡细胞、晚期凋亡细胞、活细胞及坏死细胞.华蟾素诱导细胞凋亡的最佳浓度为0.12μg/ml,最佳诱导时间为20 小时左右.结论:华蟾素具有诱导肿瘤细胞凋亡的作用.运用AO/EB染色研究细胞凋亡,具有方便、准确、可靠、便宜等优点,并能区分凋亡细胞及坏死细胞,值得推广运用.

华蟾素对肿瘤细胞周期及bcl-2蛋白表达的影响

目的 探讨华蟾素诱导肿瘤细胞凋亡的机制。方法 采用流式细胞及免疫组化的方法 ,研究华蟾素诱导肿瘤细胞凋亡与细胞周期阻断和抗凋亡基因bcl- 2表达之间的关系。结果 华蟾素能将肿瘤细胞阻断在S期 ,并能抑制bcl- 2基因的表达。结论 华蟾素诱导肿瘤细胞凋亡 ,与其能阻断细胞周期、抑制bcl-

华蟾素对乳腺癌MCF-7移植瘤裸鼠的抗肿瘤作用及机制研究

目的研究华蟾素对人乳腺癌移植瘤裸鼠的抑制作用及作用机制。方法建立MCF-7细胞裸鼠移植瘤模型,随机分为5组:模型组、顺铂阳性对照组及华蟾素3个剂量组,另设正常对照组,腹腔注射给药12 d后,取血检测血常规、肝肾功能及T淋巴细胞亚群;处死裸鼠,剥瘤称重并计算抑瘤率;取脏器,计算脏器指数,检测脾淋巴细胞增殖水平。结果华蟾素可抑制乳腺癌模型裸鼠肿瘤的生长;对荷瘤裸鼠血常规及肝肾功能无明显影响;抑制胸腺增大;促进T淋巴细胞增殖。结论华蟾素既可抑制荷瘤裸鼠肿瘤的生长,也可提高其免疫功能。

华蟾素对Hela细胞生长和小鼠脾淋巴细胞分泌IL-2的影响

目的研究华蟾素对体外培养Hels细胞生长抑制作用及其对小鼠脾淋巴细胞分泌IL-2水平的影响,探讨其抑癌作用机理、方法采用MTT法检测华蟾素对体外培养的Hela细胞生长抑制作用,体外分离纯化小鼠脾淋巴细胞,PHA-P刺激其增殖分化,并和不同浓度的华蟾素共培养,双抗体夹心ABC-ELISA法检测分泌IL-2的水平。结果华蟾素在0.0625-0.5μg/mL组间、24-96h时间组间对Hela生长抑制作用均有显著性意义(P<0.05),0.5μ/mL华蟾素与对照组间差异有显著性意义.比较0.0625-0.5μg/mL的华蟾素对小鼠脾淋巴细胞分泌lL-2水平的影响,各质量浓度组间差异均有显著性意义(P<0.05),0.25、0.5μg/mL的华蟾素与对照组间差异有显著性意义,0.0625、0.125μg/mL的华蟾素与对照组间差异无显著性意义。结论华蟾素能够抑制Hela细胞的体外生长,且能促进脾淋巴细胞分泌IL-2,从而增强T细胞免疫功能,诱导肿瘤免疫可能是华蟾素抗肿瘤的重要机制之一。

华蟾素对喉癌细胞生长及癌基因表达的影响

目的:探讨中药华蟾素(cinobufacini,CIN)对体外培养人喉癌 Hep-2细胞生长作用及癌基因表达的影响。方法:应用光镜、电镜、MTT 检测技术及 SP 免疫组化技术,对药物作用后 Hep-2细胞的数量、形态、癌基因 p53及 c-myc 蛋白的表达进行研究。结果:华蟾素对 Hep-2细胞作用在低浓度短时间时促进细胞分裂,增殖和诱导细胞分化;高浓度长时间则抑制细胞生长,促进细胞凋亡。华蟾素对 Hep-2细胞癌基因影响表现为 c-myc 蛋白表达降低,p53蛋白表达变化不明显。结论:华蟾素对喉癌细胞的生长及癌基因有抑制作用,可作为临床喉癌综合治疗的用药之一。