南方红豆杉水提物对肿瘤细胞增殖抑制作用的研究

目的研究南方红豆杉水提物对人胃腺癌细胞株SGC-7901和人乳腺癌株MCF-7增殖的抑制作用和机制。方法采用二苯基四氮唑溴盐比色法（MTT法）检测南方红豆杉水提物和紫杉醇对SGC-7901和MCF-7细胞增殖的抑制作用;光镜观察其对2种细胞形态的影响;流式细胞仪检测其对2种细胞凋亡的影响。结果南方红豆杉水提物对肿瘤细胞的增殖有抑制作用;其抑制作用机制为诱导细胞凋亡。结论南方红豆杉水提物对肿瘤细胞的增殖有抑制作用。     

紫甘蓝提取液对人乳腺癌细胞生长和迁移的抑制作用

目的:探讨紫甘蓝提取物对乳腺癌细胞MCF7生长和迁移的抑制作用。方法:应用MTT法检测不同浓度紫甘蓝提取物对MCF7增殖的抑制作用,并用倒置相差显微镜观察细胞形态,Annexin V-FITC/PI染色细胞后用流式细胞术检测不同浓度紫甘蓝提取物对MCF7凋亡的影响,采用Transwell法检测不同浓度紫甘蓝提取物对MCF7迁移的影响。采用SPSS16.0软件对实验结果进行统计学处理。结果:紫甘蓝提取物可抑制MCF7的增殖,并呈浓度依赖性和时间依赖性(P0.05),也使MCF7形态发生了异常变化,紫甘蓝提取物可诱导MCF7凋亡、抑制其迁移,且呈浓度-效应关系(P0.05)。结论:紫甘蓝提取物可抑制乳腺癌细胞MCF7生长和迁移。     

藻红蛋白诱导乳腺癌MCF-7细胞凋亡与细胞周期的关系

目的 研究藻红蛋白 (PE) 对人乳腺癌 MCF-7 细胞的增殖抑制效果和诱导凋亡作用。方法 采用 MTT 法和 SRB 法检测 PE 对人乳腺癌 MCF-7 细胞增殖的抑制作用。采用倒置显微镜、荧光显微镜 Hoechst 33258 染色观察 MCF-7 细胞凋亡细胞形态。Annexin V-FITC/PI 双荧光染色观察细胞凋亡过程中特异的磷脂酰丝氨酸 (PS) 外翻情况确定凋亡的出现。流式细胞仪观察 PE 对 MCF-7 细胞周期的影响。结果 PE 对 MCF-7 细胞有较强的生长抑制作用,并且呈一定的剂量依赖性,其 72 h 的 IC₅₀ 值为 147.82 μg/mL,GI₅₀ 值为 109.76 μg/mL。倒置显微镜下与基本贴壁的饱满、边沿清晰的对照组细胞比较,给药组细胞有明显的细胞变粗糙,贴壁细胞减少的现象,而且浓度越高,作用时间越长,悬浮细胞越多。荧光显微镜下可见各给药组 MCF-7 细胞出现不同程度深染亮点的细胞核质,染色质浓集固缩并伴有凋亡小体形成。激光共聚焦显微镜下可见给药组细胞膜被染成绿色,细胞核被染成红色。流式细胞仪显示 PE 可阻滞 MCF-7 细胞从 S 期进入 G₂/M 期,促使细胞凋亡。结论 PE 对 MCF-7 细胞有较强的抑制作用,可使其凋亡,其作用机制与周期阻滞有关。     

七叶皂苷钠通过抑制AKT,ERK上游信号SRC活性诱导人乳腺癌MCF-7细胞凋亡

探讨七叶皂苷钠对乳腺癌MCF-7细胞的凋亡诱导作用及其可能的作用机制。运用MTT法检测七叶皂苷钠对MCF-7细胞的增殖抑制作用;倒置显微镜观察细胞形态学变化;DAPI染色后在荧光显微镜下检测细胞核变化;采用Annexin V-FITC/PI流式细胞术检测细胞凋亡率;采用Western blotting检测凋亡相关蛋白(PARP,cleaved caspase-8,pro-caspase-3)和细胞存活相关信号分子(AKT,ERK)及其共同上游激酶SRC的磷酸化变化情况。结果显示,不同浓度七叶皂苷钠作用于乳腺癌MCF-7细胞后,以剂量依赖方式抑制MCF-7细胞增殖;诱导细胞凋亡(典型的凋亡形态学变化、细胞核改变和细胞凋亡率显著增加);细胞凋亡相关蛋白PARP切割增加,cleaved caspase-8表达增加,pro-caspase-3表达减少进一步验证了七叶皂苷钠的凋亡诱导作用;七叶皂苷钠显著抑制细胞存活相关信号分子(AKT,ERK)的磷酸化,其共同上游激酶SRC的活化亦显著下降。结果表明,七叶皂苷钠通过抑制SRC的活化,阻断信号向下游信号分子AKT,ERK的传递,抑制乳腺癌细胞MCF-7增殖,诱导细胞凋亡。     

蜈蚣多糖对Hela细胞的增殖抑制作用研究

目的 研究蜈蚣多糖对人宫颈癌细胞Hela细胞的抑制作用及作用机制.方法 用MTT法检测蜈蚣多糖对Hela细胞生长的抑制作用;流式细胞仪和Annexin V-FITC/PI双荧光染色分别检测蜈蚣多糖对Hela细胞周期及凋亡情况的影响.结果 在一定浓度范围内蜈蚣多糖可显著抑制Hela细胞的增殖,改变Hela细胞的细胞周期,使细胞阻滞于G2/M期,并能诱导细胞凋亡.结论 蜈蚣多糖可能通过改变Hela细胞的细胞周期,诱导细胞凋亡,从而对Hela细胞的增殖有一定的抑制作用.     

乳奥康对乳腺癌细胞株MCF-7增殖及BCL-2表达的影响

目的:探讨乳奥康对乳腺癌细胞MCF-7增殖的影响及其对BCL-2表达的影响。方法:培养乳腺癌细胞株MCF-7,采用MTT法检测乳奥康在不同浓度和不同作用时间下对乳腺癌细胞MCF-7增殖的影响;Western blotting法检测乳奥康对乳腺癌细胞BCL-2表达的影响。结果:MTT结果显示,乳奥康具有对MCF-7细胞抑制的作用,其抑制作用和浓度、作用时间呈正相关。MCF-7细胞经乳奥康作用,BCL-2的表达逐渐降低。结论:乳奥康对乳腺癌细胞MCF-7的增殖具有抑制作用,其作用机制可能是通过抑制乳腺癌细胞BCL-2的表达,从而启动炎症抑制相关机制,使肿瘤细胞发生凋亡。     

吉祥草提取物对人结肠癌HT-29细胞体外抑制作用的研究

目的 研究吉祥草提取物对人结肠癌HT-29细胞的体外增殖抑制作用和诱导凋亡作用.方法 MTT法筛选吉祥草乙醇提取物及其不同溶剂萃取部分对HT-29细胞抑制作用的活性部位;通过凋亡染色观察细胞凋亡形态;应用流式细胞术Annexin V-FITC/PI双标记法检测细胞凋亡率.结果 吉祥草正丁醇萃取物对HT-29细胞的抑制作用较强,且呈量效和时效关系,作用48 h时IC50值为66.59 mg·L-1,凋亡染色显示有典型的凋亡形态出现,流式细胞仪检测其能诱导HT-29细胞凋亡并呈剂量依赖关系.结论 吉祥草正丁醇萃取物能抑制人结肠癌HT-29细胞增殖并诱导细胞凋亡.     

雷公藤醇提物对骨髓细胞增殖和凋亡的影响

目的:观察雷公藤醇提物对大鼠骨髓细胞增殖、周期和凋亡的影响。方法:体外培养大鼠股骨骨髓细胞,经不同浓度的雷公藤醇提物作用后,采用MTT比色法检测对骨髓细胞增殖的影响;采用流式细胞仪PI单标法检测对骨髓细胞周期的影响;采用流式细胞仪Annexin V-FITC/PI双标法检测对骨髓细胞凋亡的影响。结果:雷公藤醇提物可明显抑制正常骨髓细胞增殖,并且此抑制作用呈明显的时间和剂量依赖性。细胞周期进程的结果显示,与正常对照组相比,雷公藤组G0/G1期细胞比例显著增多,S期和G2/M期细胞比例显著减少,细胞增殖指数显著降低,早期凋亡率显著升高。结论:雷公藤醇提物可明显抑制骨髓细胞增殖,使骨髓细胞阻滞于G0/G1期,并能诱导细胞凋亡。     

苦瓜皂苷对人乳腺癌MCF-7细胞增殖作用的影响

目的 研究苦瓜皂苷抑制乳腺癌MCF-7细胞的作用及其诱导凋亡的机制.方法 采用MTT法观察苦瓜皂苷对MCF-7细胞的生长抑制作用;HE染色观察皂苷作用后癌细胞的形态学变化;免疫荧光染色检测Bcl-2的阳性表达率变化;流式细胞术分析细胞周期及细胞凋亡率.结果 MTT法得到苦瓜皂苷与MCF-7细胞生长抑制率之间存在浓度依赖关系,半数致死量IC50为159.02 mg/L;HE染色观察到经苦瓜皂苷作用24h后的MCF-7细胞出现典型的凋亡形态学特征;免疫荧光检测到苦瓜皂苷作用细胞24h后,Bcl-2的阳性表达率为22.35％,远小于对照组75.61％;流式细胞术表明苦瓜皂苷处理细胞后,S期细胞数明显增加,细胞周期被阻滞在S期,随着皂苷作用时间的延长细胞凋亡率增加.结论 苦瓜皂苷可抑制体外培养MCF-7细胞乳腺癌细胞的增殖,其抑制作用与细胞凋亡机制有关.     

银杏叶提取物对乳腺癌MCF-7细胞增殖、凋亡及Caspase-3表达的影响

目的:探讨银杏叶提取物(EGB)对乳腺癌MCF-7细胞增殖、凋亡及半胱氨酸蛋白酶(Caspases)-3表达的影响.方法:将质量浓度为0,10,20,40,80,160 mg·L-1的EGB作用于体外培养的人乳腺癌MCF-7细胞上,培养24h或48 h,然后应用四甲基偶氮唑蓝染色法(MTT法)检测细胞增殖;Annexin V/PI双染色流式细胞仪检测细胞凋亡和细胞周期;酶联免疫吸附法(ELISA)检测Caspases-3蛋白表达.结果:EGB对MCF-7细胞的体外增殖具有抑制作用,量效关系显著,与对照组比较有统计学差异(P＜0.01),半数抑制浓度(IC50)为83.65 mg· L-1.经流式细胞仪检测表明,EGB能使MCF-7细胞G0-G1期逐渐增加,G2-M期和S期逐渐减少,并且随着质量浓度的增加,MCF-7细胞凋亡率明显增加(P＜0.05或P＜0.01).EGB能增强MCF-7细胞Caspases-3蛋白的表达(P＜0.05或P＜0.01),并呈浓度依赖性.结论:EGB能有效抑制人乳腺癌MCF-7细胞Caspases-3蛋白表达,诱导MCF-7细胞凋亡,抑制肿瘤细胞增殖.     

In vitro anti-breast cancer activity of ethanolic extract of Wrightia tomentosa: Role of pro-apoptotic effects of oleanolic acid and urosolic acid

Ethnopharmacological relevance

Wrightia tomentosa Roem. & Schult. (Apocynaceae) is known in the traditional medicine for anti-cancer activity along with other broad indications like snake and scorpion bites, renal complications, menstrual disorders etc. However, the anti-cancer activity of this plant or its constituents has never been studied systematically in any cancer types so far.

Aim of the study

To evaluate the anti-cancer activities of the ethanolic extract of W. tomentosa and identified constituent active molecule(s) against breast cancer.

Material and methods

Powdered leaves of W. tomentosa were extracted with ethanol. The ethanolic extract, subsequent hexane fractions and fraction F-4 of W. tomentosa were tested for its anti-proliferative and pro-apoptotic effects in breast cancer cells MCF-7 and MDA-MB-231.

Results

The ethanolic extract, subsequent hexane fractions and fraction F-4 of W. tomentosa inhibited the proliferation of human breast cancer cell lines, MCF-7 and MDA-MB-231. The fraction F-4 obtained from hexane fraction inhibited proliferation of MCF-7 and MDA-MB-231 cells in concentration and time dependent manner with IC₅₀ of 50 μg/ml and 30 μg/ml for 24 h, 28 μg/ml and 22 μg/ml for 48 h and 25 μg/ml and 20 μg/ml for 72 h respectively. The fraction F-4 induced G1 cell cycle arrest, reactive oxygen species (ROS) generation, loss of mitochondrial membrane potential and subsequent apoptosis. Apoptosis is indicated in terms of increased Bax/Bcl-2 ratio, enhanced Annexin-V positivity, caspase 8 activation and DNA fragmentation. The active molecule isolated from fraction F-4, oleanolic acid and urosolic acid inhibited cell proliferation of MCF-7 and MDA-MB-231 cells at IC₅₀ value of 7.5 μM and 7.0 μM respectively, whereas there is devoid of significant cell inhibiting activity in non-cancer originated cells, HEK-293. In both MCF-7 and MDA-MB-231, oleanolic acid and urosolic acid induced cell cycle arrest and apoptosis as indicated by significant increase in Annexin-V positive apoptotic cell counts.

Conclusion

Our results suggest that W. tomentosa extracts has significant anti-cancer activity against breast cancer cells due to induction of apoptosis pathway. Olenolic and urosolic acid are important constituent molecules in the extract responsible for anti-cancer activity of W. tomentosa.     

In vitro antiproliferative and antioxidant activities and total phenolic contents of the extracts of Melastoma malabathricum leaves

The present study aims to determine the in vitro antiproliferative and antioxidant activities of various extracts from the leaves of Melastoma malabathricum using various established in vitro assays. The aqueous extract inhibited the proliferation of Caov-3 and HL-60 cell lines, while the chloroform extract exhibited antiproliferative activity against the Caov-3, HL-60, and CEM-SS cell lines. The methanol extract demonstrated antiproliferative activity against more cell lines, including the MCF-7, HeLa, Caov-3, HL-60, CEM-SS, and MDA-MB-231 cancer cell lines. Interestingly, all extracts did not inhibit the proliferation of 3T3 cells, thus indicating their noncytotoxic properties. Unlike the chloroform extracts, the aqueous and methanol extracts of M malabathricum (20, 100, and 500 μg/ml) produced high antioxidant activity for the superoxide scavenging assay with only the 500 μg/ml aqueous and methanol extracts exhibited high activity for the 2,2-diphenyl -1-picrylhydrazyl radical scavenging assay. The total phenolic content recorded for the aqueous, methanol, and chloroform extracts were 3344.2 ± 19.1, 3055.1 ± 8.7, and 92.5 ± 7.3 mg/100 g of gallic acid, respectively. The M malabathricum leaves possessed potential antiproliferative and antioxidant activities that could be attributed to its high content of phenolic compounds.     

In vitro antiproliferative and antioxidant activities of the extracts of Muntingia calabura leaves

The in vitro antiproliferative and antioxidant activities of the aqueous, chloroform and methanol extracts of Muntingia calabura leaves were determined in the present study. Assessed using the 3,(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay, the aqueous and methanol extracts of M. calabura inhibited the proliferation of MCF-7, HeLa, HT-29, HL-60 and K-562 cancer cells while the chloroform extract only inhibited the proliferation of MCF-7, HeLa, HL-60 and K-562 cancer cells. Interestingly, all extracts of M. calabura, which failed to inhibit the MDA-MB-231 cells proliferation, did not inhibit the proliferation of 3T3 (normal) cells, indicating its safety. All extracts (20, 100 and 500 μg/ml) were found to possess antioxidant activity when tested using the DPPH radical scavenging and superoxide scavenging assays with the methanol, followed by the aqueous and chloroform, extract exhibiting the highest antioxidant activity in both assays. The total phenolic content for the aqueous, methanol and chloroform extracts were 2970.4 ± 6.6, 1279.9 ± 6.1 and 2978.1 ± 4.3 mg/100 g gallic acid, respectively. In conclusion, the M. calabura leaves possess potential antiproliferative and antioxidant activities that could be attributed to its high content of phenolic compounds, and thus, needs to be further explored.     

中药提取物乙酰胆碱酯酶抑制活性的研究

目的研究中药水提物对乙酰胆碱酯酶的抑制活性。方法选取24味中药做研究对象。应用传统的水提的方法制备中药提取物。使用改进了的Elman比色法检测24种中药的水提物对兔大脑乙酰胆碱酯酶抑制活性。结果黄柏的水提物有显著的乙酰胆碱酯酶抑制活性,在50μg/ml浓度下抑制率达75.58%,IC50值为20.03μg/ml。厚朴的水提取物在50μg/ml浓度下抑制率为43.95%,其IC50值为127.17μg/ml。益智仁的水提物有较弱的乙酰胆碱酯酶抑制活性。结论黄柏等中药提取物对乙酰胆碱酯酶具有很好的抑制活性。     

红景天对乳腺癌治疗的研究

目的：研究红景天提取物对乳腺癌MDA-MB-435细胞株增殖及血管新生相关因子的影响。方法：人乳腺癌MDA-MB-435细胞培养后,加入红景天提取物10mg／ml、20mg／ml,设置空白对照组。药物治疗24小时后,使用流式细胞仪进行分选、MTT法测定细胞增殖能力,以及VEGF表达的影响。结果：红景天处理后,MDA-MB-435细胞株的形态学特征发生显著变化,且随着药物浓度增加而愈加明显;阻滞MDA-MB-435细胞增殖周期的作用,主要阻滞于S期,该作用随着药物浓度增加亦增强;显著抑制MDA-MB-435细胞的增殖活性：处理24小时,各剂量组肿瘤细胞表达VEGF mRNA无明显差异,44小时后,给药组明显地抑制MDA-MB-435细胞表达VEGF的作用。结论：红景天提取物在体外可阻滞乳腺癌MDA-MB-435细胞增殖周期、降低乳腺癌MDA-MB-435细胞的增殖活性、减少促血管新生因子VEGF表达。     

透骨草提取物对人肝癌SMMC-7721细胞凋亡及凋亡相关因子的影响

[目的]观察透骨草提取物对人肝癌SMMC-7721细胞凋亡的影响,探讨透骨草提取物诱导SMMC-7721细胞凋亡的分子机制。[方法]流式细胞术检测透骨草提取物诱导SMMC-7721细胞凋亡的影响;免疫组化法检测透骨草提取物对SMMC-7721细胞caspase-9、-3、-7和x连锁凋亡抑制蛋白（XIAP）表达的影响。[结果]40．91μg／mL透骨草提取物处理的SMMC-7721细胞凋亡率为（26．84±1．23）％,明显高于对照组（P〈0．05）。40．9μg／mL透骨草提取物处理的SMMC-7721细胞Caspase-9、-3、-7蛋白表达明显低于与对照组（P〈0．05）,而XIAP蛋白表达明显高于对照组（P〈0．05）。瞄论]透骨草提取物能通过XIAP／Caspase信号通路抑制SMMC-7721细胞凋亡。关键词：透骨草提取物;SMMC-7721细胞;凋亡;Caspase-9中图分类号：R285．5文献标识码：A文章编号：1672—1519（2013）10—0612—03     

Water extract of Rheum officinale Baill. induces apoptosis in human lung adenocarcinoma A549 and human breast cancer MCF-7 cell lines

Ethnopharmacological relevance

Rheum officinale Baill. (Da Huang) is one of the herbs commonly used in traditional Chinese medicine formulae against cancer. The traditional decoction is similar to the water extract used in the present study.

Aim of the study

The water extract of Da Huang was investigated to see if it possesses anticancer effects through apoptotic pathways.

Materials and methods

Human lung adenocarcinoma A549 and human breast cancer MCF-7 cell lines were treated with different concentrations of Da Huang water extract at different time intervals. Growth inhibition was detected by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] and colony formation assays; apoptosis was detected by cell morphologic analysis, DNA fragmentation analysis and COMET assay.

Results

Da Huang water extract was found to have significant growth inhibitory effects on both A549 and MCF-7 cell lines with IC₅₀ values 620 ± 12.7 and 515 ± 10.1 μg/ml, respectively. Growth inhibitory effects were dose- and time-dependent. A significant decrease in cell number, DNA fragmentation and single DNA strand breakages were observed in the Da Huang water extract treated A549 and MCF-7 cells.

Conclusions

This suggests that the water extract of Da Huang exerts potential anticancer activity through growth inhibition and apoptosis on MCF-7 and A549 cells lines.     

透骨草提取物对肿瘤细胞体外增殖的影响

目的:探讨透骨草提取物对人卵巢癌SK-OV-3细胞、人乳腺癌MCF-7细胞、人宫颈癌Hela细胞、人肝癌HepG-2细胞、人肺癌A549细胞体外增殖的影响。方法:用不同浓度的透骨草提取物分别处理肿瘤细胞,采用MTT法和集落形成试验观察透骨草提取物对肿瘤细胞体外增殖的影响。结果:3.125～100μg/mL透骨草提取物处理的肿瘤细胞增殖抑制率与对照组相比显著增加(P<0.05)。3.125～100μg/mL透骨草提取物处理的肿瘤细胞集落形成率与对照组相比显著降低(P<0.05)。结论:透骨草提取物对人卵巢癌SK-OV-3细胞、人乳腺癌MCF-7细胞、人宫颈癌胞Hela细、人肝癌HepG-2细胞、人肺癌A549细胞增殖有显著的抑制作用,且呈一定的剂量依赖性。     

Differential signaling involved in Sutherlandia frutescens-induced cell death in MCF-7 and MCF-12A cells

Ethnopharmacological relevance

The scientific study of natural products traditionally used in anticancer preparations has yielded several therapeutically relevant compounds. One of these traditional preparations with potentially beneficial properties is aqueous extracts of Sutherlandia frutescens, a shrub indigenous to the Western Cape region of South Africa. The aims of this study were to evaluate in vitro efficacy of these preparations on the MCF-7 breast adenocarcinoma and MCF-12A non-tumorigenic cell lines in terms of cell proliferation, cell morphology and possible induction of cell death.

Materials and methods

Crystal violet staining was used to evaluate cell proliferation, light-and fluorescence microscopy were used to investigate both intracellular and extracellular morphological features of apoptosis and autophagy (e.g. membrane blebbing, condensed chromatin and intracellular lysosomes), while flow cytometry quantified cell cycle changes and induction of apoptosis through analysis of the flip-flop translocation of phosphatidylserine.

Results

Crystal violet staining showed a time- and dose specific response to aqueous Sutherlandia frutescens extracts, revealing exposure to 1 mg/ml aqueous extract for 48 h to be ideal for comparing the differential effects of Sutherlandia frutescens in the MCF-7 and MCF-12A cell lines. Microscopy showed distinct morphological changes with hallmarks of apoptosis being observed in both cell lines. Flow cytometry revealed a decrease in actively cycling cells in both cell lines, and a 4.36% increase in phosphatidylserine translocation in the MCF-7 cell line, indicative of apoptosis induction, while fluorescence microscopy showed evidence of the induction of autophagy.

Conclusions

Analyses revealed the carcinogenic MCF-7 cell line to be more susceptible to the cytostatic and cytotoxic effects of aqueous extracts of Sutherlandia frutescens when compared to the non-tumorigenic MCF-12A cell line, thus warranting further research into the exact cellular mechanisms involved and the possible synergistic activities of Sutherlandia frutescens ingredients.     

HPLC测定6种青海产红景天植物中红景天苷的含量

 目的 研究提取方法及HPLC测定条件以测定狭叶红景天、吉氏红景天、深红红景天、四裂红景天、大花红景天和园丛红景天中红景天苷的含量。方法 色谱柱：Spherisorb C₁₈(5 μm,4.6 mm×250 mm);流动相：甲醇-水(20∶80);流速：1.0 ml·min^-1;检测波长：223 nm。结果 甲醇回流30 min可提取完全;红景天苷线性范围为0.097～0.870 μg,平均回收率为99.6%,RSD＝1.62%。结论 该分析方法简便、灵敏、准确;红景天苷含量与红景天种类有一定的关系。