曲格列酮对人乳腺癌MDA-MB-231和MCF-7细胞株增殖影响的研究

目的:探讨过氧化物酶体增殖物激活受体γ(PPARγ)激动剂曲格列酮(TRG)对体外培养雌激素受体(ER)阴性乳腺癌MDA-MB-231细胞及ER+乳腺癌MCF-7细胞增殖的影响。方法:以不同浓度的TRG(0～50μmol/mL)分别处理体外培养的MDA-MB-231细胞和MCF-7细胞48h,MTT法检测细胞存活率;流式细胞术(FCM)检测细胞周期分布;蛋白质印迹法检测PPARγ和ER-α的表达。结果:MTT结果示TRG浓度≥5μmol/mL时MDA-MB-231细胞(P值分别为0.006、0.006、0.000和0.000)和MCF-7细胞(P值分别为0.007、0.006、0.004和0.001)的存活率降低。FCM检测结果表明,经TRG作用后肿瘤细胞主要集中在G1期。以上2种检测结果显示,MDA-MB-231细胞对TRG的敏感程度较MCF-7细胞高;蛋白质印迹法检测结果表明,TRG处理后的MDA-MB-231细胞中PPARγ蛋白的表达水平随药物浓度增加逐渐升高(相对强度分别为2.045、2.600和3.038),MCF-7细胞的ER-α表达水平则随药物浓度增加逐渐降低(相对强度分别为1.164、1.130和0.680)。结论:TRG对ER+和ER-乳腺癌细胞均有抑制作用,且ER-乳腺癌细胞对TRG更敏感。     

Estrogen- and tamoxifen-induced rearrangement of cytoskeletal and adhesion structures in breast cancer MCF-7 cells

The cytoskeleton, the shape, and the adhesion complexes of MCF-7 breast carcinoma cells have been studied by fluorescence, phase contrast, and interference reflection microscopy. Cells have been grown in media containing different concentrations of estrogen and with or without the addition of the antiestrogen tamoxifen. The pattern of actin microfilaments and keratin intermediate filaments (tonofilaments) and the distribution of adhesion areas change as a function of the estrogen concentration. When cells are cultured in estrogen-deprived medium, they appear roundish and flattened and adhere firmly to the substratum, with multiple vinculin-positive adhesion plaques at their ventral surface. Upon stimulation with estrogen, these cells display pseudopodial cytoplasmic protrusions and ruffling membranes; in interference reflection microscopy the adhesion areas are mostly localized in these projections. A rearrangement of microfilaments and of tonofilaments in the cell projections and the formation of a dense network of keratin fibers takes place. Tamoxifen affects cellular shape and cytoskeletal arrangement in a way similar to that induced by estrogen. An effect of estrogen-receptor stimulation on the adhesion structures and on the rearrangement of intermediate and actin filaments (and accordingly of the shape and internal structure of breast cancer cells) can be suggested. Such an effect might be direct or mediated through unknown mechanisms; it seems, however, to be independent of the well known estrogenic effect on cell proliferation.     

Effect of breast cyst fluid on oestrogen 17-oxidoreductase activity in MCF-7 breast cancer cells

Breast cyst fluid (BCF) was found to stimulate oestrogen 17-oxidoreductase activity in the reductive direction, i.e., conversion of oestrone (E1) to oestradiol (E2), in MCF-7 breast cancer cells. Dialysis of BCF revealed that this property of BCF was present in both dialysed BCF and dialysate, implying that both high and low mol. wt. substances were responsible for stimulating E1 to E2 conversion. Gel filtration of dialysed BCF revealed that the high mol. wt. substances responsible for the stimulation of E1 to E2 conversion had mol. wts. of approximately 11 kD and 68 kD. This property of BCF would serve to increase the concentration of E2, a steroid which may play a role in mammary carcinogenesis.     

他莫昔芬对乳腺癌MCF-7细胞侵袭能力的影响及其作用机制

目的 探讨他莫昔芬(TAM)对乳腺癌MCF-7细胞侵袭能力的影响及其作用机制.方法 采用对数生长期的乳腺癌MCF-7细胞,以无血清无酚红高糖DMEM培养基对其进行24 h的培养,然后将其分别接种并分为对照组(无血清无酚红高糖DMEM培养基)、TAM组(含有1μmol/L TAM的无血清无酚红高糖DMEM培养基),采用Western blot法、RT-PCR法检测两组MCF-7细胞基质金属蛋白酶-9(MMP-9)、MMP-2、血管内皮细胞生长因子(VEGF)蛋白及mRNA表达水平,采用Transwell法检测MCF-7细胞的侵袭能力.结果 TAM组MCF-7细胞中VEGF、MMP-9、MMP-2蛋白表达水平均明显高于对照组,VEGF、MMP-9、MMP-2 mRNA相对表达水平均明显高于对照组,差异均有统计学意义(P﹤0.01);TAM组MCF-7细胞的侵袭能力相对数明显高于对照组,差异有统计学意义(P﹤0.01).结论 TAM可能发挥类雌激素功能,促进乳腺癌MCF-7细胞侵袭能力,这与其提高MCF-7细胞中VEGF、MMP-9、MMP-2表达水平有关.     

PS341对他莫昔芬诱导乳腺癌MCF-7细胞凋亡影响及其机制的探讨

目的:研究蛋白酶体抑制剂PS341是否可增强他莫昔芬(TAM)诱导的乳腺癌细胞凋亡,并探讨可能的机制。方法:采用MTT方法测定细胞活力,采用流式细胞仪检测细胞凋亡,采用蛋白质印迹法检测Caspase-8的活化,PARP的裂解及死亡受体Fas蛋白的表达。结果:TAM以时间及浓度依赖的方式抑制乳腺癌MCF-7细胞增殖,24及48h的IC50分别为25和6μmol/L,25μmol/L的TAM处理24h可诱导21.9%的MCF-7细胞发生凋亡,10nmol/L的PS341预处理后给予25μmol/L的TAM处理24h细胞凋亡增至34.6%,PS341与TAM均可轻微上调Fas的表达水平,两药联合应用后可显著上调Fas的表达水平,并明显增强线粒体外凋亡通路的活性。结论:PS341与TAM联合应用可通过上调Fas的表达水平,增强TAM诱导的乳腺癌MCF-7细胞凋亡。     

Antiproliferative and survival properties of PMA in MCF-7 breast cancer cell

Although PKCs are assumed to be the main targets of phorbol esters (PMA), additional PMA effectors, such as chimaerins (a family of RacGTPase activating proteins) and RasGRP (exchange factor for Ras/Rap1), can counteract or strengthen the PKC pathways. In this study, we evaluated the proliferative behavior of PMA-treated MCF-7 breast cancer cell and found that: PMA induced growth arrest and inhibited cell death; PMA activated ERKs, which, in turn, induced p21; and inhibitors of ERK (PD98059) and PKC (GF109203X) prevented p21 induction and abolished the PMA survival effect. We conclude that PMA inhibits MCF-7 cell growth and simultaneously stimulates cell survival; both responses are linked to ERK-dependent and p53-independent p21 induction.     

非小细胞肺癌组织中FAK和pY397FAK的表达

目的:探讨粘着斑激酶(focaladh sionkinase,FAK)和磷酸化FAK(phosph FAK,pY397FAK)在非小细胞肺癌(no smallcelllungcancer,NSCLC)和正常支气管 黏膜上皮中的表达,及FAK在NSCLC发生 浸润和转移中的作用。方法:应用免疫组织 化学方法检测FAK和pY397FAK在60例 NSCLC和19例正常支气管黏膜上皮中的表 达,并与临床病理指标和Ki 67表达进行联 合分析。结果:FAK和pY397FAK在NSCL 中表达为83.3%(50/60)和55.0%(33 60),高于正常支气管黏膜上皮[57.9%(11 19)和21.1%(4/19)],差异有统计学意义 P=0.030、P=0.010。FAK和pY397FAK表 达升高与TNM分期及淋巴结转移有关,在 Ⅲ期和淋巴结转移组的阳性率高于Ⅰ+Ⅱ 期和无淋巴结转移组,差异有统计学意义, 均<0.05。FAK的强阳性表达率在低分化 组中(73.1%,19/26)显著高于高 中分化组 (47.1%,16/34),P=0.043。结论:FAK和 pY397FAK过表达在NSCLC发生和进展中发 挥重要作用。FAK和pY397FAK可能作为判 断NSCLC生物学行为的指标和治疗的新靶 点。     

miR-15a诱导人乳腺癌细胞MCF-7凋亡的作用机制

目的 探讨miR-15a在诱导乳腺癌细胞凋亡中的作用及机制.方法 采用定量聚合酶链反应检测人乳腺上皮细胞株MCF-10A和乳腺癌细胞株MCF-7中miR-15a的表达水平.采用软件预测miR-15a的靶点,并通过荧光素酶报告基因系统验证.将miR-15a经脂质体法转染MCF-7细胞,采用Western blot法检测Bcl-2蛋白的表达,并采用流式细胞术检测MCF-7细胞的凋亡率.结果 miR-15a在乳腺癌细胞株MCF-7中的表达明显低于乳腺上皮细胞株MCF-10A (0.253∶1,P＜0.0001).miR-15a可显著抑制抗凋亡基因Bcl-2 3′-UTR荧光素酶报告基因的表达(P＜0.05).与未转染组(对照组)相比,miR-15a转染组MCF-7细胞中Bcl-2的表达水平显著下降,凋亡明显增加(P＜0.05).结论 miR-15a 作为一个潜在的抑癌基因,可通过靶向于Bcl-2诱导乳腺癌细胞MCF-7发生凋亡,其可为乳腺癌的临床治疗提供新的靶点和理论依据.     

Effect of RNA interference of iASPP on the apoptosis in MCF-7 breast cancer cells

ASPP family is proved to be apoptotic specific regulators of p53. Among them, iASPP acts as an inhibitor of p53. To investigate the effect of the iASPP RNAi on the apoptosis of breast cancer cell MCF-7, we transfected the recombinant plasmid PGCsilencer H1/Neo/GFP/RNAi into MCF-7. The iASPP expression was analyzed by RT-PCR and Western blot. The cell apoptosis was detected by FCM. The results show that the expression of iASPP is descended and the apoptosis rate is increased after transfection. Therefore, we conclude that inhibition of expression of iASPP may resume the ability of p53 to induce apoptosis in MCF-7 cells.     

弗林蛋白酶抑制剂对乳腺癌MCF-7细胞生长和迁移的影响

目的 探讨乳腺癌转移的机制,为深入研究乳腺癌发生、发展机制提供理论基础.方法 应用不同浓度的弗林蛋白酶（Furin）抑制剂α1-PDX处理乳腺癌MCF-7细胞.用四甲基偶氮唑蓝（MTT）和克隆形成实验检测Furin抑制剂对MCF-7细胞增殖和克隆形成的影响.单层细胞迁移实验和Transwell实验检测MCF-7细胞迁移和浸润能力.Hoechst 33342/PI双染法检测细胞凋亡.酶联免疫吸附法检测细胞培养液中基质金属蛋白酶2（MMP-2）和MMP-9蛋白水平.Western blot检测细胞迁移相关蛋白MT1-MMP、血管内皮生长因子（VEGF）-C和VEGF-D水平.结果 不同浓度α1-PDX作用MCF-7细胞48 h以上时,细胞的生长受到抑制,集落形成降低,细胞凋亡率升高.但在低浓度情况下对细胞迁移和侵袭起抑制作用.α1-PDX降低了细胞内MT1-MMP、VEGF-C、VEGF-D的表达,同时细胞培养液上清中MMP-2和MMP-9浓度也低于对照组.结论 Furin抑制剂通过抑制乳腺癌MCF-7细胞MMP及VEGF表达抑制肿瘤的迁移能力.     

Specific induction of pp125 focal adhesion kinase in human breast cancer

The pp125 focal adhesion kinase (FAK) is involved in integrin-mediated cell signalling and overexpressed in a variety of solid tumours. Focal adhesion kinase expression has been correlated to invasion and metastasis, but the data on breast cancer are inconclusive. We analysed FAK mRNA, protein levels and expression patterns in primary breast cancer and normal breast tissue. FAK expression on the functional protein level and mRNA was determined in 55 matched pairs of breast cancer and corresponding normal tissue by Western blot, immunohistochemistry and RT-PCR. Using a score ranging from 0 to +5 for Western blots, we determined in normal breast tissue a score of 1.51+/-0.84 (mean+/-standard deviation), which was strongly induced to 2.91 (+/-1.22) in breast cancers (P<0.001). Overall, 45 out of 55 tissue pairs (81.8%) showed this upregulation of FAK protein in tumours in comparison to normal tissue. Immunohistochemistry confirmed these findings with a significant higher score for tumours vs physiological tissue (1.0+/-0.63 vs 2.27+/-0.91; P=0.001). Interestingly, no overall significant difference in the mRNA levels (P=0.359) was observed. In conclusion, expression levels of the FAK protein are specifically upregulated in breast cancer in comparison to matched normal breast tissue supporting its pivotal role in neoplastic signal transduction and representing a potential marker for malignant transformation.     

Estrogen regulates Ah responsiveness in MCF-7 breast cancer cells

Cytochrome P450 (CYP)1A1 and CYP1B1, which are under the regulatory control of the aryl hydrocarbon (Ah) receptor (AhR), catalyze the metabolic activation of numerous procarcinogens and the hydroxylation of 17beta-estradiol (E2) at the C-2 and C-4 positions, respectively. There is evidence of cross-talk between estrogen receptor alpha (ERalpha)- and AhR-mediated signaling in breast and endometrial cells. To further examine these interactions, we investigated the short- and long-term effects of E2 exposure on Ah responsiveness in MCF-7 human breast cancer cells. Short-term exposure to 1 nM E2 elevated the ratio of the 4- to 2-hydroxylation pathways of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced E2 metabolism and the ratio of the induced CYP1B1 to CYP1A1 mRNA levels, as determined by real-time PCR. Cells maintained long-term (9-12 months) in low-E2 medium progressively lost Ah responsiveness, as indicated by diminished rates of TCDD-induced E2 metabolism and ethoxyresorufin O-deethylase activity, and the reduced expression of the CYP1A1 and CYP1B1 mRNAs and proteins levels. These E2-deprived cells showed elevated levels of ERalpha mRNA, depressed levels of AhR mRNA, and unchanged levels of the AhR nuclear translocator mRNA. Transient transfection studies using a CYP1B1-promoter-luciferase reporter construct showed that reduced CYP1B1 promoter activity in E2-deprived cells could be restored by co-transfection with an AhR expression construct, indicating that AhR expression was limiting in these cells. The reduced Ah responsiveness of E2-deprived cells was reversed by culture for four passages in medium supplemented with 1 nM E2; ERalpha and AhR mRNAs returned to near-normal levels and the inducibility of the CYP1A1 and CYP1B1 mRNAs, proteins, and E2 metabolic activities by TCDD was restored. These studies indicate that the continued presence of estrogen is required to maintain high levels of AhR expression and inducibility of the procarcinogen-bioactivating enzymes, CYP1A1 and CYP1B1, in MCF-7 cells.     

洛铂对ＭＣＦ-７细胞凋亡及ＭＴＤＨ基因表达的影响

目的　研究洛铂对人乳腺癌ＭＣＦ-７细胞凋亡的作用并探讨其对转移基因ＭＴＤＨ表达的影响。方法　ＭＴＴ比色法检测洛铂对ＭＣＦ-７细胞株的增殖抑制作用,流式细胞仪检测洛铂对ＭＣＦ-７细胞的凋亡率,ＲＴ-ＰＣＲ法检测洛铂对乳腺癌转移相关基因ＭＴＤＨ表达的影响。结果　５～４０ｍｇ／Ｌ洛铂对ＭＣＦ-７细胞增殖均具有明显抑制作用,呈药物浓度 时间依赖性（Ｐ＜０.０５）;５ｍｇ／Ｌ、１０ｍｇ／Ｌ、１５ｍｇ／Ｌ洛铂作用于ＭＣＦ-７细胞４８ｈ后凋亡率分别为１９.０８％、２４.２３％和６５.７１％。洛铂可以抑制ＭＴＤＨ基因表达,当浓度≥１０ｍｇ／Ｌ时,ＭＴＤＨｍＲＮＡ表达呈阴性。结论　洛铂可抑制ＭＣＦ-７细胞的增殖并可诱导其发生凋亡,并可能通过抑制ＭＴＤＨ基因的表达抑制乳腺癌细胞的侵袭转移。     

碧萝芷对乳腺癌MCF-7细胞增殖、迁移和转移的影响

目的探讨抗氧化剂碧萝芷对乳腺癌MCF-7细胞的抑制作用,并对观察碧萝芷对MCF-7细胞的转移和迁移功能的影响。 方法用10、20、40 μg/ml的碧萝芷处理乳腺癌MCF-7细胞作为实验组,空白对照组只加入DMEM培养基(无血清), MTT法在处理细胞后24、48、72 h于波长490 nm处测定各组吸光度值;用流式细胞仪检测细胞凋亡率;用β-半乳糖苷酶染色法检测MCF-7细胞衰老率;用Transwell小室法分别检测各浓度碧萝芷对乳腺癌MCF-7细胞迁移和侵袭能力的影响;用Western blot法分别检测乳腺癌MCF-7细胞衰老相关蛋白P53、P21、P16、P27、E2F1以及基质金属蛋白酶(matrix metalloproteinase, MMP)-2、MMP-9的表达。细胞迁移、侵袭数目及各种蛋白的表达水平比较采用单因素方差分析,吸光度值比较采用重复测量的方差分析,两两比较采用LSD法。 结果MTT结果显示,在处理乳腺癌MCF-7细胞24、48、72 h后,各组吸光度值比较,差异有统计学意义(F＝149.439, P<0.001),在不同时间点之间比较,差异有统计学意义(F＝27.922,P<0.001);分组与时间点之间存在交互作用(F＝18.466, P<0.001)。流式细胞仪检测结果显示：空白对照组及10、20、40 μg/ml碧萝芷组的细胞凋亡率分别为(2.36±0.27)%、(6.44±1.43)%、(7.52±2.09)%和(11.68±1.65)%,差异有统计学意义(F＝19.143, P<0.001)。β-半乳糖苷酶染色法结果显示：空白对照组和10、20、40 μg/ml碧萝芷组的细胞衰老率分别为(5.35±1.32)%、(20.08±2.14)%、(40.55±4.61)%和(59.26±4.10)%,差异有统计学意义(F＝150.150, P<0.001)。Transwell小室结果显示：空白对照组及10、20、40 μg/ml碧萝芷组的迁移细胞数目分别为(41±5)、(27±2)、(19±1)、(11±2)个,差异有统计学意义(F＝59.330, P<0.001);侵袭转移的细胞数目分别为(24±4)、(17±1)、(12±2)、(7±2)个,差异也有统计学意义(F＝26.230, P<0.001)。Western blot结果显示：碧萝芷可上调P53、P21、P16、P27蛋白表达(F＝263.905、424.937、217.515、391.115,P均<0.001),下调E2F1蛋白的表达(F＝64.003,P<0.001);同时,各组MCF-7细胞中的MMP-2及MMP-9蛋白表达水平明显降低(F＝44.104、45.594, P均<0.001)。 结论碧萝芷可能是以促衰老的方式抑制乳腺癌MCF-7细胞的生长,且能抑制细胞的迁移和转移。     

脂质体介导c-myc反义寡核苷酸对乳腺癌MCF-7细胞生物学行为的影响

目的研究c-myc反义寡核苷酸(ASODN)经脂质体LipfectAMINETM(LR)介导转染对MCF-7细胞中c-myc蛋白表达及细胞增殖、凋亡的影响。方法用四甲基偶氮唑盐法(MTT法)评价脂质体介导c-myc ASODN对细胞增殖的影响;免疫细胞化学ABC方法检测转染前后c-myc蛋白表达;流式细胞仪(FCM)定量分析细胞凋亡。结果ASODN/LR转染与正义、错义寡核苷酸相比,显著抑制MCF-7细胞增殖(P<0.01)。FCM分析结果显示,转染后可见明显细胞凋亡峰,72h相点细胞凋亡率达(28.76±2.09)%。免疫细胞化学显示c-myc蛋白水平明显降低,阳性表达率为(21.40±1.16)%。结论LR介导c-myc ASODN能明显抑制MCF-7细胞增殖、诱导细胞凋亡和下调c-myc蛋白水平。     

Effect of hyperosmolality on alkaline phosphatase and stress-response protein 27 of MCF-7 breast cancer cells

Summary MCF-7, a continuous cell line derived from a human breast carcinoma, exhibits very low alkaline phosphatase (ALP) activity. The enzyme is heat-stable and is inhibited by L-phenylalanine and L-phenylalanylgly-cylglycine, but not by L-homoarginine, 1-bromotetramisole, or levamisole. These data indicate that MCF-7 produces term-placental ALP, the oncodevelopmental enzyme form inappropriately expressed by a variety of human tumors. In contrast to human cancer cells that produce this enzyme monophenotypically, ALP activity of MCF-7 cells is not significantly increased by glucocorticoids or sodium butyrate. By comparison, exposure to hyperosmolality causes a striking increase in enzyme activity. Cycloheximide blocks this effect. The results obtained with cell-free assays were confirmed by cytochemical and immunocytochemical assays on whole cells. Because some of the agents tested in the enzyme modulation experiments affect cell proliferation, their possible effect on two stress-response proteins (srp 27 and srp 72) was also examined; specific immunocytochemical assays were used. These tests revealed that neither protein is affected by glucocorticoids; that sodium butyrate has no effect on srp 27, but alters the intracellular distribution of srp 72; and that hyperosmolality, while not significantly affecting srp 72, causes an increase in srp 27.     

人乳腺癌MCF-7细胞中SP亚群细胞的分离及其生物学行为研究

目的:了解肿瘤干细胞的生物学行为及相关的调控机制.方法:采用干细胞对Hoechst33342染料外排的特性进行MCF-7 细胞系肿瘤干细胞的流式分选.并对其增殖、自我更新、分化能力等生物学行为进行研究.还利用流式细胞仪、免疫荧光技术、荧光定量PCR等技术进行了与干细胞生物学行为密切相关的Wnt信号转导通路中相关分子的表达特征研究.结果:MCF-7细胞系中包含SP亚群(side population);且SP细胞具有分化潜能,高增殖能力,及自我更新特性;而且Wnt通路的各种关键分子在该细胞系SP亚群细胞中呈活化状态.结论:MCF-7细胞系的SP亚群可以代表该细胞系的肿瘤干细胞,Wnt通路在该肿瘤细胞系肿瘤干细胞的自我更新等其它生物学行为的调控中起重要作用.     

Vascular endothelial growth factor receptor-3 and focal adhesion kinase bind and suppress apoptosis in breast cancer cells

Focal adhesion kinase (FAK) and vascular endothelial growth factor receptor-3 (VEGFR-3) are protein tyrosine kinases that are overexpressed in human cancer and play an important role in survival signaling. In addition to its involvement with cell survival, VEGFR-3 is a primary factor in lymphatic angiogenesis. Because FAK function is regulated by its COOH terminus (FAK-CD), we used FAK-CD as a target to identify binding partners. We isolated a peptide from a phage library that bound to FAK-CD, specifically the focal adhesion targeting domain of FAK and was homologous to VEGFR-3, suggesting these two tyrosine kinases physically interact. We have also shown that VEGFR-3 is overexpressed in human breast tumors and cancer cell lines. For the first time, we have shown the physical association of FAK and VEGFR-3. The association between the NH(2) terminus of VEGFR-3, containing the peptide identified by phage display, and the COOH terminus of FAK was detected by in vitro and in vivo binding studies. We then coupled a 12-amino-acid VEGFR-3 peptide, AV3, to a TAT cellular penetration sequence and showed that AV3 and not control-scrambled peptide caused specific displacement of FAK from the focal adhesions and affected colocalization of FAK and VEGFR-3. In addition, AV3 peptide decreased proliferation and caused cell detachment and apoptosis in breast cancer cell lines but not in normal breast cells. Thus, the FAK/VEGFR-3 interaction may have a potential use to develop novel molecular therapeutics to target the signaling between FAK and VEGFR-3 in human tumors.     

Src激酶抑制剂PP2在乳腺癌MCF-7细胞转移中的作用和机制

  目的  探讨Src激酶抑制剂PP2在乳腺癌MCF-7细胞转移中的作用和机制。  方法  乳腺癌MCF-7细胞经PP2作用48 h后, 检测肿瘤细胞体外粘附、侵袭能力的改变, 细胞周期的改变, Western blot及Real-time PCR试验检测肿瘤细胞转移相关基因表达的变化, 报告基因试验检测AP-1和NF-κB启动子活性的变化。  结果  PP2可抑制MCF-7细胞中Src激酶活性, 2.5μM和5μMPP2作用后, 肿瘤细胞粘附能力下降63.4%和34.7%;侵袭能力下降44.3%和20.2%;细胞周期显著阻滞在G₀/G₁期; CD44、MMP-2/9以及p-β-catenin表达显著下降, E-cadherin表达显著升高; AP-1启动子活性下降64.5%和37.9%, NF-κB启动子活性下降55.7%和31.8%。  结论  Src激酶与乳腺癌MCF-7细胞肿瘤转移能力密切相关, 阻断Src激酶活性可抑制肿瘤细胞转移能力。      

ON123300对ER阳性人乳腺癌MCF-7细胞的影响及机制研究

目的 初步探讨新型多靶点激酶抑制剂ON123300对ER阳性、HER2阴性的人乳腺癌MCF-7细胞增殖、周期、凋亡的影响及机制。方法 取处于对数生长期的MCF-7细胞,分为DMSO对照组、Palbocilib处理组及ON123300处理组,给药干预48 h后观察细胞生长状态,CCK-8法检测各组MCF-7细胞增殖抑制情况,流式细胞术(Flow cytometry,FCM)检测细胞周期和凋亡,Western blot检测cyclinD1、CDK4、pRb1、survivin和PI3K的表达。结果 光学显微镜下可见DMSO对照组MCF-7细胞贴壁良好,增殖迅速;Palbocilib处理组的MCF-7细胞增殖出现轻微抑制和少量细胞脱落,未见细胞形态改变;ON123300处理组MCF-7细胞增殖明显受抑制,数量明显减少,细胞形态改变,出现脱落现象。细胞抑制率随药物浓度增加而增加,呈现剂量依赖效应。ON123300处理组的MCF-7细胞明显阻滞在G₁/S期,并伴有细胞凋亡增加,差异有统计学意义(P＜0.05)。ON123300处理组cyclinD1、CDK4、pRb1、survivin、PI3K的蛋白表达量随药物浓度增高均逐渐降低,差异具有统计学意义(P＜0.05)。结论 ON123300对人乳腺癌细胞MCF-7有显著的增殖抑制、促进细胞周期阻滞和凋亡的作用,其机制可能与survivin/cyclinD1/CDK4/Rb1/和PI3K信号通路的抑制相关。