Molecular mechanism about lymphogenous metastasis of hepatocarcinoma cells in mice

AIM: To investigate the correlation between lymphogenous metastasis and matrix metalloproteinases (MMPs) activity and the expression of Fas ligand of tumor cells in lymph nodes. METHODS: Fifty-six inbred 615-mice were equally divided into 2 groups and inoculated with Hca-F and Hca-P cells. Their lymph node metastatic rates were examined. Growth fraction of lymphocytes in host lymph nodes was detected by flow cytometry. The Hca-F and Hca-P cells were cultured with extract of lymph node, liver or spleen. The quantity of MMPs in these supernatants was examined by zymographic analysis. The expression of Fas ligand, PCNA, Bcl-2 protein of Hca-F and Hca-P cells in the mice were examined by immunohistochemistry. The apoptosis signals of macro-phages in lymph nodes were observed with in situ DNA fragmentation. RESULTS: On the 28th day post-inoculation, the lymph node metastatic rate of HcaF was 80%(16/20), whereas that of Hca-P was 25%(5/20). The growth fraction of lymphocytes was as follows: in the Hca-F cells, the proliferating peak of lymphocytes appeared on the 14th day post inoculation and then decreased rapidly, while in HcaP cells, the peak appeared on the 7th day post inoculation and then kept at a high level. With the extract of lymph node, the quantity of the MMP-9 activity increased (P【0.01) and active MMP-9 and MMP-2 were produced by both Hca-F and Hca-P tumor cells, which did not produce MMPs without the extract of lymph node or with the extracts of the liver and spleen. The expression of Fas Ligand of Hca-F cells was stronger than that of Hca-P cells (P 【0.01). The expressions of PCNA and Bcl-2 protein of Hca-F cells in the tumors of inoculated area were the same as that of Hca-P cells. In situ DNA fragmentation showed that the positive signals of macrophages were around Hca-F cells. CONCLUSION: Secretion of MMPs which was associated with metastatic ability of Hca-F and Hca-P tumor cells depends on the environment of lymph nodes. The increased expression of Fas ligand protein of Hca-F tumor cells with high lymphogenous metastatic potential in lymph nodes may help tumor cells escape from being killed by host lymphocytes.

Effect of 5-Aza-2'-deoxycytidine on the P16 tumor suppressor gene in hepatocellular carcinoma cell line HepG2

INTRODUCTIONHepatocellular carcinoma (HCC) is one of the mostcommon human malignancies worldwide[1,2], and isclosely associated with infection of HBV and HCVand contamination of aflatoxin B1[3-6]. Althoughthe molecular mechanisms of hepatocarcinogenesisremain poorly understood, an increasing number ofgenetic abnormalities have been recognized[7-10],for example, the p16 gene[11,12] the p53gene[13-18], the E-cadherin gene[19], and the c-mycgene[20].

Growth of Circulating Tumor Cell-Derived Colonies from Peripheral Blood of Melanoma Patients: Preliminary Characterization of Colony Composition

Circulating tumor cells (CTC) are rarely detected in the blood of cancer patients, even though they are a direct harbinger of eventual patient demise. We developed an innovative CTC culture technology to allow more sensitive isolation, expansion, and characterization of viable colonies from patient blood. In this assay, the entire leukocyte fraction from 10 ml of anticoagulated patient blood is placed into culture medium without any pre-selection. After 16 days in culture, CTC derived colonies are counted. As a proof-of-principle, blood samples from 58 Stage IIa-IV melanoma patients were tested. Ninety percent of these samples grew colonies. The colony numbers ranged from 0 - 308 (mean 63 ± 9.5 SEM). Ten normal volunteers had virtually no growth (mean 0.5 ± 1.4 colonies). Colonies were harvested using a micropipette for characterization. Tumor-cell containing spheroids were embedded in paraffin, sectioned, and stained with melanoma-specific mAb for histologic characterization. MITF proved to be the most consistent immunostain that identified melanoma cells in these colonies. A host-cell component in colonies was also identified using CD68 and CD43 mAb staining. Following enzymatic dissociation of colonies, a variety of immunostains were tested. Papanicolau staining proved most useful for identifying the abnormal nuclei of tumor cells. Flow cytometry could readily distinguish host and tumor cell populations based on DNA content and forward/side scatter in dissociated colonies. The stem cell marker ALDH1A1 associated with the aneuploid population, but CD45 was expressed on both diploid and aneuploid cells. The ability to repeatedly isolate CTC derived colonies from cancer patient blood samples opens the door to a novel type of long-term clinical monitoring. This novel CTC culture technology may prove useful to perform molecular characterization, assessment of treatment response, and testing of drug sensitivity and resistance in patients during treatment.

Cell cycle analysis by cyclin E+A/DNA multiparameter flow cytometry in exponential growth MOLT-4 cells

Objective To establish a new method for analyzing the cell cycle in tumor which is a kind of cell cycle disease.Methods Mixtures of cyclin E and cyclin A antibodies were incubated with fixed MOLT-4 cells, and measured by flow cytometry.Results We developed a cyclin E+A/DNA flow cytometry analysis method, which may distinguish G0, early G1, late G1, S, G2 and M phase cells, rather than three phases in the DNA content histogram.Conclusion Cyclin E+A/DNA multiparameter flow cytometry can simultaneously differentiate in the same sample six cell groups: G0, early G1, late G1, S, G2 and M phase cells. It performed better than any other cell cycle analysis methods that we have used and has a definite cell biology foundation.

Apoptosis susceptibility of tumor cells to arsenic trioxide and the inherent cellular level of reactive oxygen species

OBJECTIVE: To explore the association of inherent cellular reactive oxygen species (ROS) levels with susceptibility of the tumor cells to apoptosis induction by arsenic trioxide (As(2)O(3)). METHODS: Low concentration (2 micromol/L) of As(2)O(3) was administered to two cultured leukemic cell lines, NB4 and U937, and two esophageal carcinoma cell lines, EC1.71 (also named EC/CUHK1) and EC1867, to confirm the difference in apoptosis susceptibility of NB4 versus U937 and of EC1.71 versus EC1867. Dihydrogenrhodamine 123 (DHR123), used as a ROS capture agent, was incubated with cells in the absence of As(2)O(3). Fluorescence intensity of rhodamine 123, the product of cellular oxidation of DHR123, was detected by flow cytometry and ROS was measured. RESULTS: Low concentration of As(2)O(3) induced apoptosis was more likely to occur in NB4 and EC1.71 cells than in U937 and EC1867 cells, or NB4 was more sensitive than U937, and EC1.71 more sensitive than EC1867 to As(2)O(3). The inherent cellular ROS level is higher in NB4 than in U937, and also higher in EC1.71 than in EC1867. CONCLUSIONS: The difference in cellular ROS level is positively associated with cellular susceptibility to apoptosis induction by As(2)O(3). The inherent ROS level might be important in defining apoptotic susceptibility to As(2)O(3).

咖啡因促受照射肝癌细胞株MHCC97H凋亡的实验研究

目的观察咖啡因促受照射MHCC97H细胞株凋亡的作用及机制。方法通过流式细胞仪分析不同条件下细胞凋亡比例和周期分布,应用Western Blotting动态观察磷酸化CDC2 Tyr15的表达量,利用细胞克隆集落试验证明咖啡因的放疗增敏效果。结果MHCC97H细胞照射后48小时,0、8、16、24 Gy组的凋亡率分别为5.7%、12.0%、19.4%、21.7%;咖啡因促进凋亡,2.5 mmol/L咖啡因组的凋亡比例在照射16 Gy 48小时左右由对照组的19.4%升至32.3%,并且随着时间的推移凋亡比例逐渐增加。辐射引起细胞G2期阻滞,0、8、16、24 Gy组的G2期比例分别为13.9%、25.6%、41.4%、51.6%,G2期阻滞的程度与剂量呈正相关;而咖啡因可去除由辐射引起的G2期阻滞,2.5 mmol/L咖啡因组的G2期细胞比例在照射12小时左右由对照组的37.6%降为29.6%。MHCC97H细胞照射后,G2期阻滞程度与磷酸化CDC2 Tyr15的表达量一致;咖啡因则抑制磷酸化CDC2 Tyr15的表达量。细胞克隆集落试验显示,咖啡因可降低放射后MHCC97H细胞的存活分数,放疗增敏比(SER)为1.28。结论咖啡因通过增加CDC2 Tyr15的脱磷酸化,去除由辐射引起的G2期阻滞,从而促进凋亡,达到放疗增敏的效果。

细胞凋亡检测用于肿瘤细胞株对化疗敏感性的研究

目的 评价 Annexin- V荧光标记 FACScan法、TU NEL法细胞凋亡检测在肿瘤细胞株化疗药物敏感性研究中的应用。方法 将 DDP、MMC、5 - FU、EPI按血浆峰浓度 (PPC)、1/10 PPC、1/5 PPC、5 PPC、10 PPC与结肠腺癌L o Vo和 L s- 174- t细胞温育 2 4及 48h,用 FACScan 法、TU NEL法检测细胞凋亡 ,用 MTT比色法检测细胞生长抑制率 ,并提取 DNA进行琼脂糖凝胶电泳。结果 L o Vo与L s- 174- t细胞经化疗药物诱导 48h后 ,FACScan检测的细胞凋亡率在 PPC时最高 ,TU NEL 法检测的凋亡率与 MTT法检的细胞增殖抑制率呈正的直线相关 (P<0 .0 5 )。结论 Annexin- V荧光标记 FACScan法检测细胞凋亡既能优选出敏感的药物 ,又能寻找合适的药物浓度 ,是目前优选有效化疗药物种类及剂量的一个可探索。

全反式维甲酸和5-Fu对胃癌细胞端粒酶活性和细胞生长的影响

目的观察 ATRA,5-Fu 单独和联合应用对体外培养的胃癌细胞端粒酶活性及细胞生长的影响.方法分别用 ATRA,5-Fu 单独和联合处理胃癌 MGC-803细胞,采用 MTT 法测定细胞活力,采用端粒重复序列扩增法测定端粒酶活性.结果胃癌细胞活力随 ATRA,5-Fu 浓度增高、作用时间的延长其细胞活力逐渐下降,ATRA d1,d3,d5的 IG_(50)分别为>40μmol/L,(20～40)μmol/L,(10～20)μmol/L;5-Fu d1,d3,d 5的 IC_(50)分别为>25/μmol/L,(2～5)/μmol/L,(2～5)μmol/L;40μmol/L ATRA,5μmol/L 5-Fu 及40μmol/LATRA+5μmol/L 5-Fu 处理胃癌细胞3 d 其细胞活力分别为46%,47%,8%(P<0.01),端粒酶活性分别为45.68%(P<0.01),100.00%,46.10%(P<0.01).结论 ATRA,5-Fu 均能抑制胃癌细胞的生长,其抑制作用具有时间和浓度依赖性,且二者合用具有协同抑制作用;ATRA能抑制胃癌细胞端粒酶活性,而5-Fu 对胃癌细胞端粒酶活性无影响,二者合用对胃癌细胞端粒酶活性无协同抑制作用.抑制端粒酶活性可能是 ATRA 抗癌机制之一.

丹参酮Ⅱ_A对人肝癌BEL-7402细胞生长的影响及其机制

目的研究丹参酮ⅡA对体外培养的人肝癌细胞系BEL-7402细胞生长的影响及其作用机制.方法0～10μg/ml丹参酮ⅡA作用人肝癌BEL-7402细胞72 h后,用倒置相差显微镜观察丹参酮ⅡA对BEL-7402细胞的生长抑制作用;荧光显微镜、透射电镜、DNA琼脂糖凝胶电泳检测细胞凋亡;流式细胞术检测不同浓度丹参酮作用后细胞凋亡率的变化.结果肝癌细胞经丹参酮ⅡA作用后,细胞生长明显受到抑制;荧光显微镜、透射电镜观察发现细胞皱缩、核质浓缩、核碎裂、细胞起泡及凋亡小体形成等凋亡特征性形态改变;琼脂糖凝胶电泳观察到DNA'梯状带';流式细胞仪定量分析示浓度为0.5、1.0、2.0、5.0、和10.0μg/ml的丹参酮ⅡA处理肝癌细胞72 h后,细胞凋亡率分别为(20.78±2.17)%、(24.64±2.07)%、(31.47±3.86)%、(43.65±4.04)%和(52.36±3.75)%,与对照组[(2.37±0.29)%]比较,均有显著性差异.结论丹参酮ⅡA能抑制体外培养的人肝癌细胞系BEL-7402细胞的生长,其机制可能为诱导细胞凋亡.

DHA复合物对鼠移植瘤细胞和T淋巴细胞细胞周期及凋亡的影响

目的 研究DHA复合物的抗癌机理。方法 用流式细胞术研究了DHA复合物对荷瘤鼠H2 2 细胞和T淋巴细胞细胞周期及凋亡的影响。结果 ( 1)与阴性对照组相比 ,DHA复合物中、高剂量组G0 G1期H2 2 癌细胞百分比明显增加 (P <0 .0 1) ;DHA复合物低、中、高剂量组G0 G1期T细胞百分比明显减小 (P <0 .0 1)。 ( 2 )与阴性对照组相比 ,DHA复合物低、中、高剂量组S期H2 2 癌细胞百分比明显减小 (P <0 .0 1) ,S期T细胞百分比明显增加 (P <0 .0 1)。 ( 3)与阴性对照组相比 ,DHA复合物中剂量组G2 M期H2 2 癌细胞百分比显著减少 (P <0 .0 1) ,DHA复合物低、高剂量组G2 M期H2 2 癌细胞以及DHA复合物低、中、高剂量组G2 M期T细胞百分比均显著升高 (P <0 .0 1)。 ( 4 )与阴性对照组相比 ,DHA复合物低、中、高剂量组H2 2 癌细胞增殖指数 (PI)明显下降 (P <0 .0 1) ,T细胞增殖指数明显升高 (P <0 .0 1)。 ( 5 )与阴性对照组相比 ,DHA复合物低、中、高剂量组H2 2 癌细胞及DHA复合物中、高剂量组T细胞凋亡率明显升高 (P <0 .0 1)。结论 DHA复合物可抑制H2 2 癌细胞的增殖 ,促进T淋巴细胞增殖 ,同时DHA复合物还可促进H2 2 细胞。

瑞香狼毒诱导HL-60细胞凋亡和调节SGC-7901细胞bcl-2蛋白表达(英文)

目的 探索瑞香狼素 (SC)抗肿瘤机制。方法 以 HL- 6 0和 SGC- 790 1为靶细胞 ,用 MTT比色法测定细胞增殖抑制 ,荧光显微镜观察凋亡细胞的形态学改变 ,DNA电泳和流式细胞仪检测DNA断裂 ,免疫组化检测 bcl- 2蛋白表达。结果 含 SC药物血清处理细胞 48h后 ,HL- 6 0细胞增殖呈剂量依赖性抑制 ,并表现出典型的凋亡细胞形态学改变及 DNA断裂 :即染色体聚集、核固缩、断裂及阶梯状 DNA电泳条带 ,G1 期前细胞从 11.7%增至 5 7.4% ;而 SGC- 790 1细胞 bcl- 2蛋白表达率从 78.3%下降到 32 .9%。结论 SC可诱导肿瘤细胞凋亡 ,降低 bcl- 2蛋白表达。

胃癌细胞系SGC-7901肿瘤干细胞样细胞的分离和鉴定

目的探讨人胃癌细胞系SGC-7901中鉴定胃癌干细胞样细胞。方法采用无血清培养基培养胃癌细胞系SGC-7901,得到悬浮生长状态的细胞球;流式细胞术分别检测贴壁胃癌细胞和悬浮细胞球中Side Population(SP)侧群细胞的含量;NOD/SCID鼠体内异位移植成瘤实验检测成瘤能力;基因芯片技术筛选悬浮细胞球与贴壁细胞差异表达的基因;实时荧光定量PCR验证基因芯片结果。结果 (8.00±0.54)%SGC-7901细胞在无血清培养基中能增殖并形成可连续传代的细胞球;悬浮细胞球中SP细胞百分含量高达(12.34±1.01)%,贴壁细胞情况中SP仅占(0.62±0.05)%;NOD/SCID小鼠成瘤实验显示,悬浮球细胞组致瘤能力高于贴壁细胞组;基因芯片结果显示,肿瘤球高表达与干细胞相关的基因ABCG2、HES1和KRT153。实时荧光定量PCR方法检测结果与基因芯片结果基本一致。结论胃癌细胞系SGC-7901中存在肿瘤干细胞样细胞。

Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97

AIM: To establish clone cells with different metastatic potential for the study of metastasis-related mechanisms. METHODS: Cloning procedure was performed on parental hepatocellular carcinoma (HCC) cell line MHCC97, and biological characteristics of the target clones selected by in vivo screening were studied. RESULTS: Two clones with high (MHCC97-H) and low (MHCC97-L) metastatic potential were isolated from the parent cell line. Compared with MHCC97-L, MHCC97-H had smaller cell size (average cell diameter 43 microm vs 50 microm) and faster in vitro and in vivo growth rate (tumor cell doubling time was 34.2h vs 60.0h). The main ranges of chromosomes were 55-58 in MHCC97-H and 57-62 in MHCC97-L. Boyden chamber in vitro invasion assay demonstrated that the number of penetrating cells through the artificial basement membrane was (37.5 +/- 11.0) cells/field for MHCC97-H vs (17.7 +/- 6.3)/field for MHCC97-L. The proportions of cells in G0-G1 phase, S phase, and G2-M phase for MHCC97-H/MHCC97-L were 0.56/0.65, 0.28/0.25 and 0.16/0.10, respectively, as measured by flow cytometry. The serum AFP levels in nude mice 5wk after orthotopic implantation of tumor tissue were (246 +/- 66) microg.L(-1) for MHCC97-H and (91 +/- 66) microg.L(-1) for MHCC97-L. The pulmonary metastatic rate was 100% (10/10) vs 40% (4/10). CONCLUSION: Two clones of the same genetic background but with different biological behaviors were established, which could be valuable models for investigation on HCC metastasis.

奥沙利铂对肝癌细胞HepG2细胞周期的影响

目的:探讨奥沙利铂(Oxaliplatin,L-OHP)对人肝癌细胞株HepG2周期的影响及其相关机制,为其应用于原发性肝细胞癌临床治疗提供理论依据。方法:MTT方法检测L-OHP对肝癌细胞HepG2生长的抑制作用;流式细胞术(FCM)检测L-OHP诱导HepG2细胞周期阻滞的作用;RTPCR和Western blot方法观察L-OHP对细胞周期调节因子CyclinD1、CDK2、CDK4及p16、p21、p53表达的影响。结果:MTT结果显示L-OHP对HepG2细胞增殖抑制率随药物浓度增加及作用时间延长有升高趋势,呈时间剂量依赖性。L-OHP能够诱导HepG2细胞周期阻滞于S期,并可下调CDK4和CyclinD1蛋白的表达,上调p21,p53蛋白的表达,CDK2和p16表达无明显变化。结论:L-OHP可能通过影响CDK4、CyclinD1及p21的活性使HepG2细胞阻滞在S期,从而抑制肝癌细胞HepG2的增殖。

流式细胞术对不同肿瘤细胞表面所表达血小板免疫相关抗原的检测

目的：研究不同肿瘤细胞表面血小板免疫相关抗原的表达。方法：利用流式细胞术观察了ＰＧＣＬ３、ＰＡａ、ＰＧ－３、ＰＣ－３Ｍ、ＭＧＣ８０３、ＥＳＣＬ、ＢｅＬ、ＴＣＴ、ＫＢ、Ａ２７８０、ＣＣＡ８０１共１１种人肿瘤细胞，其中包括二对高低不同转移能力的人肺癌（ＰＧＣＬ３和ＰＡａ）和人前列腺癌（ＰＧ－３和ＰＧ－３Ｍ）细胞阳性表达不同血小板免疫相关抗原的细胞数及其肿瘤细胞表面抗原表达的平均荧光强度。结果：１１种人肿瘤细胞膜表面均有ＣＤ９、ＣＤ６３、ＣＤ４２ａ和ＴＳＰ，等血小板免疫相关抗原不同程度的表达，在ＭＧＣ８０３细胞膜表面发现有ＣＬ８６较强的表达，ＣＤ４１、ＣＤ４２ｂ、ＣＤ６１和ＣＤ６２均未发现表达。１１种人肿瘤细胞系中ＣＤ９＋、ＣＤ４２ａ＋、ＣＤ６３＋、ＴＳＰ＋细胞所占的比例各不相同，不同肿瘤细胞系每个抗原阳性细胞抗原表达的荧光强度也不同。与高转移ＰＧＣＩ３和ＰＧ－３Ｍ细胞相比，ＣＤ９在低转移细胞系ＰＡａ和ＰＧ－３上有较高的表达，ＣＤ４２ａ、ＴＳＰ有相对低的表达。ＣＤ４２ａ＋和ＴＳＰ＋细胞数和表达强度在高转移ＰＧＣＬ３和ＰＧ－３Ｍ细胞系均要高于低转移的ＰＡａ和ＰＧ－３；

p16^(INK4α)重表达对人食管癌细胞系EC109生长的抑制作用

目的探讨用腺病毒介导野生型的 p16基因在食管癌细胞系表达,研究重表达野生型 p16基凶对食管癌细胞系 EC109生长的抑制作用,为寻找食管癌致癌机制的研究及其基因治疗提供理论基础.方法用基因重组技术,将 pcDNA3-p16中的野生型 p16基因用 Kpn I/Bam H (?)进行双酶切,克隆入腺病毒表达载体pAdCMV 中,将重组质粒 pad-CMV-p16与腺病毒质粒 JM17用Lipofectamime 2000共转染293细胞,产生重组腺病毒.用重组腺病毒感染人食管癌细胞系 EC109,在感染后的不同时段,用免疫荧光、打点杂交方法检测 p16基因在细胞中的表达.用MTT 法及流式细胞仪观察 p16基因的表达对食管癌细胞株生长的影响.结果经酶切鉴定野生型 p16基因克隆入腺病毒表达载体中,与 JM17共转染293细胞后,可产生具有感染活性的重组腺病毒,用重组腺病毒感染食管癌细胞株 EC109细胞后,可抑制食管癌细胞的生长,同对照组相比其最大抑制率可达52.7%.Multipcycle 分析软件分析对细胞周期影响表明,处于 G_0～G_1期的细胞为41%～63%.感染后的细胞经 Dot blot 和 Westernblot 杂交证实,有外源的 p16mRNA 及蛋白的表达.结论重组腺病毒可将野生型 p16基因导入人食管癌细胞系EC109中,野生型 p16基因在 p16基因功能缺失的细胞中重表达能抑制癌细胞恶性生长.p16基因功能缺失是食管癌致癌因素之一.

ω-3多不饱和脂肪酸对胃癌BGC-823细胞凋亡的影响

目的利用人胃癌BGC-823细胞体外培养实验,观察不同含量ω-3多不饱和脂肪酸合成的二十碳五烯酸(eicosapentaenoic acid,EPA)和二十二碳六烯酸(docosahexaenoc acid,DHA)对肿瘤细胞凋亡的影响。方法采用MTT法、凋亡形态学检查和流式细胞术检测肿瘤细胞凋亡情况。结果30和45 mg/L的EPA或DHA可显著降低肿瘤细胞存活率(P<0.01),透射电镜可见细胞凋亡现象,流式细胞检测bcl-2和Gα基因蛋白表达明显下降(P<0.05,P<0.01)。结论ω-3多不饱和脂肪酸可诱导人胃癌BGC-823细胞凋亡,细胞增殖受到抑制。

Relationship between Fas/ FasL expression and apoptosis of colon adenocarcinoma cell lines

INTRODUCTIONFas/ FasL system has been identified as a keymediator of apoptosis in tumor cells[1-4]. Theoccurrence and development of neoplasm are closelyrelated to apoptosis[5-7] Most chemotherapeuticdrugs kill cancer cells mainly by inducingapoptosis[8-14].'

Studies on mechanism of Sialy Lewis-X antigen in liver metastases of human colorectal carcinoma

INTRODUCTIONSialyl Lewis-X antigen ,correlated with carcinoma, is a group of carbohydrate antigen containing oligosaccharide expressed of embryonic tisue and glycoproteins on cell surface of embryonic tissue[1].The SLeX antigen located on cell surface is synthesized principally by two enzymes ,al ,3fucosyltransfrease and a2, 3sialyctransferase.In adults ,SLeX antigen is expressed principally on the surfaces of granulocytic cells and some tumor cells .

三苯氧胺诱导乳腺癌细胞凋亡过程中ERα和SMRT的表达变化

目的:观察三苯氧胺(tamoxifen,TAM)作用前后乳腺癌细胞株T-47D和MDA-MB-231中雌激素受体α(estrogen receptor alpha,ERα)、共抑制因子SMRT的表达变化,寻找更佳的指导乳腺癌内分泌治疗指标。方法:采用四氮甲基唑蓝(MTT)观察乳腺癌细胞T-47D和MDA-MB-231的生存状况,以筛选最佳的药物作用浓度和时间。流式细胞仪法(FCM法)检测细胞凋亡。细胞免疫组化法及Western blot法观察TAM作用前后细胞中ER-α、SMRT的表达情况。结果:MTT法测得经0.10 mmol/L TAM作用48 h后T-47D和MDA-MB-231细胞增殖率下降,与对照组相比有显著性差异(P<0.05),特别对T-47D效果更强。FCM法检测两种细胞均出现低于G1期DNA含量的亚二倍体凋亡峰,与MDA-MB-231相比,T-47D细胞株凋亡率更高。细胞免疫组化法及Western blot法检测T-47D细胞,ERα、SMRT表达阳性,ERα阳性细胞比例减少,SMRT阳性细胞比例增多;MDA-MB-231细胞SMRT阴性,出现少量ER阳性及SMRT阳性细胞。结论:TAM可以通过诱导细胞凋亡途径抑制乳腺癌细胞增殖,对ERα阳性细胞T47D效果更为显著。