黄荆子乙酸乙酯提取物诱导人宫颈癌Hela细胞凋亡

[目的]探讨黄荆子乙酸乙酯提取物（EVn-50）体外诱导人宫颈癌Hela细胞的凋亡作用。[方法]体外培养Hela细胞,不同浓度EVn-50（1.0、10.0、100.0μg/ml）作用于Hela细胞。采用PI染色FCM检测EVn-50诱导Hela细胞的凋亡作用;采用间接免疫荧光标记FCM检测凋亡相关蛋白Bcl-2、Bax、Caspase-3的表达。[结果]EVn-501.0、10.0、100.0μg/ml处理Hela细胞48h后,细胞的凋亡率分别为1.06%、8.80%及20.90%,呈浓度依赖性（P〈0.05）。不同浓度EVn-50作用Hela细胞48h,细胞中Bax、Caspase-3蛋白表达逐渐升高,Bcl-2蛋白表达逐渐降低（P〈0.05）。[结论]EVn-50具有诱导Hela细胞凋亡作用,其作用可能与改变凋亡相关蛋白Bax、Caspase-3及Bcl-2的表达有关。     

扇贝提取物诱导Hela细胞凋亡的研究

目的探讨扇贝提取物对体外培养Hela细胞有无凋亡诱导作用。方法体外培养的Hela细胞经不同浓度的扇贝提取物处理24h,经Hoechst 33258(8g/L)染色进行形态学观察、琼脂糖凝胶电泳及流式细胞术(FCM)检测细胞。结果加扇贝提取物处理后,荧光显微镜下观察发现培养细胞中出现核固缩、凋亡小体,琼脂糖凝胶电泳可见DNA ladder条带,流式细胞仪检测有凋亡峰。结论扇贝提取物可诱导体外培养Hela细胞凋亡。     

人参皂苷Rg3对人宫颈癌Hela细胞的诱导凋亡作用

目的:探讨人参皂苷Rg3对人宫颈癌Hela细胞的诱导凋亡作用。方法:培养人宫颈癌Hela细胞,不同浓度Rg3处理,HE染色观察细胞形态学改变,CCK-8、流式细胞仪检测细胞的凋亡。结果:Rg3作用后Hela细胞出现了典型的凋亡形态学变化,包括细胞皱缩、细胞核内染色体浓缩。10、20、40、80μg/ml人参皂苷Rg3处理48小时后Hela细胞的平均生存率为76.3%、57.1%、50.0%、29.8%;流式细胞术(FCM)显示肿瘤细胞凋亡率为16.4%、37.0%、50.0%、69.8%,表明肿瘤细胞在人参皂苷Rg3诱导下发生凋亡。结论:Rg3能抑制Hela细胞增殖,诱导其凋亡,其诱导凋亡作用呈剂量依赖性。     

多柔比星对人宫颈癌Hela细胞凋亡的形态学观察

目的:从形态学方面探讨多柔比星影响Hela细胞生长的作用机制.方法:采用MTT法观察多柔比星对人宫颈癌Hela细胞生长的影响,并采用倒置显微镜、荧光显微镜、透射电镜和激光扫描共聚焦显微镜观察Hela细胞形态变化.结果:MTT实验发现,2 mg/L的多柔比星对Hela细胞的生长有显著的抑制作用,培养4 d后,其生长抑制率达到55.51%,而对L929细胞的生长抑制率仅为18.51%.形态学观察结果表明,多柔比星作用4 d后的Hela细胞呈典型的凋亡形态学改变.结论:2 mg/L多柔比星是通过诱导Hela细胞凋亡而抑制其生长的.     

细胞同步化对肿瘤坏死因子诱导Hela细胞凋亡的影响

目的 研究细胞周期时相对肿瘤坏死因子（TNF）诱导Hela细胞凋亡的影响。方法 应用胸腺嘧啶核苷（TdR）双阻断法将培养的Hela细胞同步化,采用MTT法、流式细胞术和荧光染色,分析同步化的Hela细胞和正常培养的Hela细胞对TNF（加放线菌酮增效）诱导凋亡的敏感性。结果 同步化Hela细胞较正常培养的Hela细胞对TNF诱导调亡的敏感性增强。结论 TNF诱导Hela细胞凋亡与细胞周期有关。     

8-Br-cAMP诱导人宫颈癌Hela细胞凋亡的实验研究

目的探讨以8-Br-cAMP体外诱导Hela细胞凋亡中,相关基因及蛋白质表达与凋亡之间的关系,为使用无毒性8-Br-cAMP治疗宫颈癌提供依据.方法采用TUNEL法检测凋亡细胞,采用原位杂交和完整细胞原位斑点印迹技术检测凋亡相关基因及蛋白质的表达.结果8-Br-cAMP试验组细胞凋亡率为(40.0±1.32)%,对照组(5.2±0.74)%;8-Br-cAMP可上调wp53,iNOS基因表达,下调mp53,bc1-2,c-myc基因表达,可增强iNOS和FasL的酶活性,降低Fas免疫反应性.以上各结果,试验组与对照组相比P＜0.01.结论8-Br-cAMP能诱导Hela细胞凋亡,可作为一种治疗宫颈癌的新途径.     

细胞周期对肿瘤坏死因子诱导Hela 细胞凋亡的影响

 目的　研究细胞周期对肿瘤坏死因子 ( TNF)诱导 Hela细胞凋亡的影响。方法　通过胸腺嘧啶核苷酸 ( Td R)阻断法阻滞 Hela细胞的细胞周期 ,以 MTT法、流式细胞术和荧光染色分析 Td R阻滞细胞和周期化的 Hela细胞对 TNF诱导凋亡的敏感性。结果　Td R阻滞细胞周期较周期化的 Hela细胞对 TNF诱导的凋亡的敏感性降低。结论　揭示 TNF诱导 Hela细胞凋亡与细胞周期有关.     

热疗对人宫颈癌Hela细胞凋亡的影响

目的:观察不同加温温度对人宫颈癌Hela细胞凋亡的影响.方法:人宫颈癌Hela细胞株常规方法培养,采用水浴加热法(温度为41℃、42.5℃、43.5℃、)对细胞进行加温处理,处理后继续培养24h.用流式细胞仪检测细胞凋亡,单细胞凝胶电泳(single cell gel electrophoresis,SCGE)法检测DNA受损状态.结果:随着温度的增加细胞凋亡率增加,42.5℃、43.5℃加温1h后细胞凋亡率最高分别为30.7%和34.6,坏死细胞分别为13.2%和29.6%.单细胞凝胶电泳发现42.5℃加温1h后40.0%的细胞有DNA损伤,43.5℃加温1h后80.0%以上的细胞DNA损伤,而41℃加温处理1h后仅有20.0%细胞DNA受损.结论:单独加温处理1h可诱导细胞凋亡,并导致细胞DNA损伤.     

缺氧诱导宫颈癌Hela细胞的生物学行为改变

[目的]研究缺氧引起的糖酵解改变对宫颈癌Hela细胞生物学行为的影响。[方法]CoCl2化学诱导宫颈癌Hela细胞缺氧,将实验对象分为常氧组和缺氧组。用MTT法、软琼脂克隆形成实验、流式细胞仪、跨膜侵袭实验等方法观察两组细胞的增殖、克隆、凋亡和侵袭能力;用酶比色法检测细胞培养液上清中HKⅡ和乳酸值。免疫细胞化学法和Western blot法分别检测两组HIF-1α、GLUT1的表达,RT-PCR检测两组HIF-1α、GLUT1及HKⅡ基因表达水平。[结果]缺氧组与常氧组相比,缺氧组细胞生长增殖活跃、凋亡率下降、跨膜细胞数增加,差异有显著性意义(P<0.05)。缺氧组中HKⅡ和乳酸值较常氧组明显升高(P<0.05)。缺氧诱导后,HIF-1α基因表达水平无明显变化(P>0.05),蛋白表达水平明显升高(P<0.05),GLUT1基因和蛋白在缺氧诱导后表达水平都明显升高(P<0.05)。[结论]氧微环境下宫颈癌Hela细胞增殖、抗凋亡、侵袭能力加强,可能与缺氧诱导后HIF-1α蛋白不宜降解,细胞糖酵解能力加强有关。     

Apoptosis of Hela cell induced by Celastrus orbiculatus Thunb extract and primary mechanisms

Objective  

The apoptosis of Hela cells induced by ethyl acetate extract and n-butanol extract of Celastrus orbiculatus Thunb was studied in order to assess its antitumor effect.     

顺铂诱导人卵巢癌细胞系HO-8910细胞凋亡

[目的]探讨顺铂诱导卵巢癌细胞凋亡作用的机制.[方法]应用不同浓度顺铂处理卵巢癌HO-8910细胞,于不同时间收集细胞,分别进行倒置显微镜观察活细胞,Gimsa染色观察细胞形态改变并计数凋亡率,琼脂糖凝胶电泳观察DNA降解情况,SP免疫组化染色观察凋亡过程中p53蛋白的表达情况.部分涂片还使用了TUNEL染色计数凋亡细胞,并与Gimsa染色计数的凋亡细胞数作比较.[结果]在顺铂作用下,卵巢癌HO-8910细胞呈现典型的凋亡细胞形态学改变,如细胞核固缩、染色体凝聚、有凋亡小体形成,细胞DNA电泳呈梯形带,凋亡率及凋亡相关蛋白p53表达阳性率随着药物作用时间延长及药物浓度增大而升高,且二者呈正相关.TUNEL染色计数的凋亡细胞个数明显高于Gimsa染色.[结论]顺铂可以通过诱导卵巢癌细胞系HO-8910细胞凋亡而发挥其抗肿瘤功效,且为p53依赖性细胞凋亡过程.     

Momordica cochinchinensis Aril Extract Induced Apoptosis in Human MCF-7 Breast Cancer Cells

Momordica cochinchinensis Spreng (MC) has been used in traditional medicine due to its high carotenoidcontent. The objective of this study was to investigate mechanisms underlying apoptotic effects of MC on humanMCF-7 breast cancer cells. A lycopene-enriched aril extract of MC (AE) showed cytotoxicity and antiestrogenicityto MCF-7 cells. On DAPI staining, AE induced cell shrinkage and chromatin condensation were evident. Withflow cytometric analysis, AE increased the percentage of cells in an early apoptosis stage when compared withthe control group. RT-PCR analysis showed AE to significantly increase the expression of the proapoptotic baxgene without effect on expression of the anti-apoptotic bcl-2 gene. Moreover, AE enhanced caspase 6, 8 and 9activity. Taken together, we conclude that AE of MC fruit has anticancer effects on human MCF-7 breast cancercells by induction of cell apoptosis via both intrinsic and extrinsic pathways of signaling     

Silibinin Enhances Ultraviolet B-Induced Apoptosis in MCF-7 Human Breast Cancer Cells

Purpose

Chemotherapies for breast cancer generally have strong cellular cytotoxicity and severe side effects. Thus, significant emphasis has been placed on combinations of naturally occurring chemopreventive agents. Silibinin is a major bioactive flavonolignan extracted from milk thistle with chemopreventive activity in various organs including the skin, prostate, and breast. However, the mechanism underlying the inhibitory action of silibinin in breast cancer has not been completely elucidated. Therefore, we investigated the effect of silibinin in MCF-7 human breast cancer cells and determined whether silibinin enhances ultraviolet (UV) B-induced apoptosis.

Methods

The effects of silibinin on MCF-7 cell viability were determined using the MTT assay. The effect of silibinin on PARP cleavage, as the hallmark of apoptotic cell death, and p53 protein expression in MCF-7 cells was analyzed using Western blot. The effect of silibinin on UVB-induced apoptosis in MCF-7 cells was analyzed by flow cytometry.

Results

A dose- and time-dependent reduction in viability was observed in MCF-7 cells treated with silibinin. Silibinin strongly induced apoptotic cell death in MCF-7 cells, and induction of apoptosis was associated with increased p53 expression. Moreover, silibinin enhanced UVB-induced apoptosis in MCF-7 cells.

Conclusion

Silibinin induced a loss of cell viability and apoptotic cell death in MCF-7 cells. Furthermore, the combination of silibinin and UVB resulted in an additive effect on apoptosis in MCF-7 cells. These results suggest that silibinin might be an important supplemental agent for treating patients with breast cancer.     

康莱特注射液诱发肾癌细胞凋亡及p53,bcl—2表达的研究

目的;探讨康莱特注射液抗肿瘤的作用机制。方法：利用ＭＴＴ法分析康莱特肾癌细胞的抑制作用,末端脱氧核苷酰转移酶法检测细胞凋亡,免疫组织化学法分析ｐ５３和ｂｃｌ－２基因表达的影响。结果：康莱特抑制肾癌细胞的ＩＣ５０为１９．３１μｌ／ｍｌ,５μｌ／ｍｌ,和１０μｌ／ｍｌ康莱特注射液具有诱发细胞凋亡的作用,细胞凋亡分别为３１．３０％和８９．７６％,康莱特浓度继续增加时,细胞凋亡的数量反而减少,１５μdispla     

卡铂复合液诱导胃癌细胞凋亡和p53蛋白表达的研究

作者比较了卡铂液和卡铂复合液对胃癌细胞株（ＮＫＭ－２５）细胞凋亡和ｐ５３蛋白表达的影响，研究表明随着卡铂液作用时间的延长，胃癌细胞凋亡数量和ｐ５３蛋白表达明显增加，免疫组织化学研究亦显示卡铂液作用后可以使胃癌细胞ｐ５３蛋白的表达增强。卡铂复合液对胃癌细胞凋亡数量和ｐ５３蛋白表达的影响与卡铂液的作用相比未见显著性差异（Ｐ＞０．０５）。研究提示：①卡铂液诱导胃癌细胞凋亡与ｐ５３蛋白的表达密切相关，可能为ｐ５３蛋白依赖性；②卡铂复合液保持了卡铂液所具有的对胃癌细胞凋亡的诱导作用。结合作者的前期研究，提示采用卡铂复合液腹腔内化疗，腹腔粘连和腹膜纤维化程度明显减轻，并发症少，是治疗胃肠道癌肿等的一种安全、有效的区域化疗方法。     

p38 MAPK Signaling Mediates Mitochondrial Apoptosis in Cancer Cells Induced by Oleanolic Acid

Oleanolic acid (OA) is a nutritional component widely distributed in various vegetables. Although it has beenwell recognized for decades that OA exerts certain anti-tumor activity by inducing mitochondria-dependentapoptosis, it is still unclear that what molecular signaling is responsible for this effect. In this study, we employedcancer cell lines, A549, BXPC-3, PANC-1 and U2OS to elucidate the molecular mechanisms underlying OA antitumoractivity. We found that activation of MAPK pathways, including p-38 MAPK, JNK and ERK, was triggeredby OA in both a dose and time-dependent fashion in all the tested cancer cells. Activation was accompaniedby cleavage of caspases and PARP as well as cytochrome C release. SB203580 (p38 MAPK inhibitor), but notSP600125 (JNK inhibitor) and U0126 (ERK inhibitor), rescued the pro-apoptotic effect of OA on A549 and BXPC-3 cells. OA induced p38 MAPK activation promoted mitochondrial translocation of Bax and Bim, and inhibitedBcl-2 function by enhancing their phosphorylation. OA can induce reactive oxygen species (ROS)-dependentASK1 activation, and this event was indispensable for p38 MAPK-dependent apoptosis in cancer cells. In vivo,p38 MAPK knockdown A549 tumors proved resistant to the growth-inhibitory effect of OA. Collectively, weelucidated that activation of ROS/ASK1/p38 MAPK pathways is responsible for the apoptosis stimulated byOA in cancer cells. Our finding can contribute to a better understanding of molecular mechanisms underlyingthe antitumor activity of nutritional components.     

Mechanisms of Gambogic Acid-Induced Apoptosis in Non-Small Cell Lung Cancer Cells in Relation to Transferrin Receptors

Abstract

Gambogic acid (GA) is one of the important active ingredients of gamboge. Our study examined the expression of transferrin receptors (TFR) on the cell surface of human lung SPC-A1 and SK-MES-1 cells and measured their GA-induced apoptosis rate. The results showed that SPC-A1 cells with a higher TFR expression were more sensitive at the same GA concentrations. To examine its distribution in cultured cells and study the mechanisms of apoptosis, we labeled GA with a ¹²⁵I tracer and examined the expression of apoptosis-related proteins. We found that GA uptake into SPC-A1 cells was higher than into SK-MES-1 cells; apoptosis-related proteins Caspase 2, Caspase 9, Caspase 10, Bax and p53 were involved in GA-induced apoptosis. We conclude that GA has an apoptosis-promoting effect on non small cell lung cancer cells. In clinical practice, the histopathological quantitation of TFR expression levels in tumor tissues may become a predictor of the sensitivity of patients' tumors to GA treatment.