A novel taspine derivative,HMQ1611, suppresses adhesion,migration and invasion of ZR-75-30 human breast cancer cells

Background

Taspine was screened for the first time from Radix et Rhizoma leonticis (Hong Mao Qi in Chinese) using cell membrane chromatography in our laboratory. Its anticancer and antiangiogenic properties were demonstrated, and it could serve as a lead compound in anticancer agent development. Here, we investigated the role of one of the derivatives, HMQ1611, with increased activity and solubility, on the regulation of breast cancer cell ZR-75-30 adhesion, migration and invasion.

Methods

The effect of HMQ1611 on adhesion, invasion and migration of human breast cancer cells ZR-75-30 was examined. The migration and invasive potential of ZR-75-30 cells were examined by wound-healing assays and matrigel invasion chamber assays. The adhesion to type IV collagen and laminin were evaluated by MTT assay. The expression and proteinase activity of two matrix metalloproteinases (MMPs), matrix metalloproteinases 2 (MMP-2) and matrix metalloproteinases 9 (MMP-9), were analyzed by Western blot analysis and gelatin zymography, respectively.

Results

HMQ1611 effectively inhibited ZR-75-30 cell invasion and significantly suppressed adhesion to type IV collagen and laminin-coated substrate in a dose-dependent manner. Western blot and gelatin zymography analysis showed that HMQ1611 significantly inhibited the expression and secretion of MMP-2 and MMP-9 in ZR-75-30 cells. Additionally, treatment of ZR-75-30 cells with HMQ1611 downregulated the expression of MMP-2 and MMP-9.

Conclusions

HMQ1611 had potential to suppress the adhesion, migration and invasion of ZR-75-30 cancer cells, and it could serve as a potential novel therapeutic candidate for the treatment of metastatic breast cancer.     

Steroidal Saponins from Paris polyphylla Suppress Adhesion,Migration and Invasion of Human Lung Cancer A549 Cells Via Down-Regulating MMP-2 and MMP-9

Background: Tumor metastases are the main reasons for oncotherapy failure. Paris polyphylla (Chinese name:Chonglou) has traditionally been used for its anti-cancer actions. In this article, we focus on the regulation ofhuman lung cancer A549 cell metastases and invasion by Paris polyphylla steroidal saponins (PPSS). Materialsand Methods: Cell viability was evaluated in A549 cells by MTT assay. Effects of PPSS on invasion and migrationwere investigated by wound-healing and matrigel invasion chamber assays. Adhesion to type IV collagen andlaminin was evaluated by MTT assay. Expression and protease activity of two matrix metalloproteinases (MMPs),MMP-2 and MMP-9, were analyzed by Western blotting and gelatin zymography, respectively. Results: PPSSexerted growth inhibitory effects on A549 cells, and effectively inhibited A549 cell adhesion, migration andinvasion in a concentration-dependent manner. Western blotting and gelatin zymography analysis revealed thatPPSS inhibited the expression and secretion of MMP-2 and MMP-9 in A549 cells. Conclusions: PPSS has thepotential to suppress the migration, adhesion and invasion of A549 cells. PPSS could be a potential candidatefor interventions against lung cancer metastases.     

基质金属蛋白酶-9反义寡核苷酸对卵巢癌细胞侵袭黏附能力的影响

目的 观察基质金属蛋白酶-9（MMP-9）反义寡核苷酸（ASODN）转染对卵巢癌细胞体外侵袭黏附行为的影响,并探讨其作用机制。方法 以Lipofectinmin介导的MMP-9反义寡核苷酸转染至经纤黏连蛋白诱导MMP-9表达的卵巢癌细胞株HO-8910PM,利用RT—PCR、Western blot及明胶酶谱法检测转染寡核苷酸后HO-8910PM细胞MMPO的mRNA、蛋白表达及酶活性的变化;通过细胞体外侵袭、迁移实验和黏附实验,检测细胞侵袭黏附能力的变化。结果 卵巢癌细胞HO-8910PM转染MMP-9反义寡核苷酸后,MMP-9的mRNA及蛋白的表达受到抑制,抑制率分别为34．8％和42．5％,与对照组比较,差异有统计学意义（P〈0．05）;明胶酶活性也受到了抑制。反义寡核苷酸的转染降低了肿瘤细胞体外侵袭、迁移和黏附能力,侵袭和迁移抑制率分别为22．4％和24．8％,在60min和90min黏附抑制率分别为49．8％和38．3％。结论 MMP-9反义寡核苷酸可抑制卵巢癌细胞的侵袭黏附能力,MMP-9有可能成为抗卵巢癌侵袭转移的分子靶点。     

Soybean saponin inhibits tumor cell metastasis by modulating expressions of MMP-2, MMP-9 and TIMP- 2

The antimetastatic properties of soybean saponin were investigated by evaluating matrix metalloproteinase (MMP-2 and MMP-9) production in HT-1080 cells. The mRNA expression levels of MMP-2 and MMP-9 were determined by RT-PCR analysis. The levels of secreted MMP-2, MMP-9 and tissue inhibitor of metalloproteinase-2 (TIMP-2) were determined by gelatin zymography and/or ELISA. The invasion of a Matrigel-coated membrane by human fibrosarcoma HT-1080 and HT-29 colon cancer cells was quantitatively assessed by counting the migrated cells. The treatment of HT-1080 cells with soybean saponin inhibited the mRNA expression of and reduced the amounts of secreted MMP-2 and MMP-9, whereas it increased the amount of secreted TIMP-2 dose-dependently. Soybean saponin significantly inhibited the invasion of HT-1080 cells through a Matrigel-coated membrane. The antimetastatic properties by soybean saponin were further confirmed by in vivo mice experiment via the tail vein injection of CT-26 colon cancer cells after feeding the mice the dietary soybean saponin. The incidence of metastatic tumor colonization of lungs of mice moderately decreased 2 weeks after the tail vein injection of CT-26 cells. Our current data support the notion that soybean saponin inhibits tumor cell metastasis by suppressing MMP-2 and MMP-9 productions, and stimulating TIMP-2 secretion, thereby suggesting that soybean saponin has a chemopreventive property against cancer metastasis.     

基质金属蛋白酶抑制剂LY52对人卵巢上皮癌细胞SKOV3中MMP-2,MMP-9表达及其侵袭转移能力的抑制作用

背景与目的：研究表明,肿瘤细胞MMP-2,MMP-9表达升高与其侵袭和转移能力有密切关系。因而,研究设计新的MMPs特异性抑制剂得到广泛重视。根据MMP-2,MMP-9结构特点,本实验设计合成全新结构的N1-取代咖啡酰吡咯烷类MMPs抑制剂——LY52,并观察该化合物对MMP-2,MMP-9表达的抑制作用及对人卵巢粘液囊腺上皮癌细胞SKOV3侵袭转移能力的影响。方法：MTT和细胞克隆法检测LY52对细胞生长抑制作用;明胶酶直接降解琥珀酰明胶法测定LY52抑酶作用活性：SDS-PAGE zymography分析对SKOV3细胞MMP-2,MMP-9表达的抑制作用;MTT法测定细胞与FN,LN和matfigel的粘附能力：Transwell小室法观察化合物对SKOV3细胞侵袭人工基底膜的抑制作用。结果：直接与细胞接触,LY52具有较弱的肿瘤细胞生长抑制作用。LY52直接抑制明胶酶活性,抑制作用IC50为11．9μg／ml。同时,LY52抑制SKOV3细胞MMP-2,MMP-9表达。以0．1、1、10、100、1000μg／ml剂量与SKOV3细胞孵育24h．对培养上清液MMP-2表达的抑制率为10．66％-31．47％：对MMP-9表达的抑制率为22．56％-56．71％。按上述剂量孵育1h。对SKOV3细胞与FN,LN和matfigel粘附能力的最大抑制率分别为29．79％,48．94％,50．92％。LY52显著抑制SKOV3细胞穿过人工基底膜能力,按上述剂量,LY52与细胞孵育24h,抑制率分别为7．5％,42．07％,66．54％,72．15％。82．84％。结论：LY52通过抑制SKOV3细胞MMP-2,MMP-9的表达．降低癌细胞侵袭转移能力。     

Flavonoid baicalein suppresses adhesion,migration and invasion of MDA-MB-231 human breast cancer cells

Baicalein is a widely used Chinese herbal medicine that has been used historically in anti-inflammatory and anti-cancer therapy. However, the molecular mechanism of its anti-cancer activity remains poorly understood and warrants further investigations. The purpose of this study is to verify the activity of baicalein to inhibit the invasion of MDA-MB-231 human breast cancer cells. The results indicated that baicalein suppressed MDA-MB-231 cell adhesion to fibronectin-coated substrate, wound healing migration and invasion through the Matrigel in a concentration-dependent manner. Western blot and gelatin zymography analysis showed that baicalein significantly inhibited the expression and secretion of matrix metalloproteinases 2/9 (MMP-2/9) in MDA-MB-231 cells. Additionally, treatment of MDA-MB-231 cells with baicalein down-regulated the expression of MMP-2/9 involved mitogen-activated protein kinases (MAPK) signaling pathway. Taken together, baicalein had potential to suppress the adhesion, migration and invasion of MDA-MB-231 cancer cells in vitro and it could serve as a promising drug for the treatment of cancer metastasis.     

肿瘤和间质细胞相互作用对肺癌MMP-2和MMP-9表达的影响

 目的 研究三维立体培养中肺癌H460细胞、胚肺成纤维细胞和单核细胞相互作用对MMP-2、MMP-9 表达的影响。 方法 活性胶原三维立体培养分为：H460细胞和胚肺成纤维细胞共同培养组;H460细胞和单核细胞共同培养组;胚肺成纤维细胞和单核细胞共同培养组;H460细胞、胚肺成纤维细胞和单核细胞共同培养组4组。明胶酶谱法测定各组培养基中MMP-2、MMP-9活性。 结果 H460细胞和胚肺成纤维细胞、胚肺成纤维细胞和单核细胞共同培养时MMP-2酶原和活化酶活性和表达均增强,H460细胞和单核细胞共同培养时MMP-2酶原和MMP-9活性和表达增强,H460细胞、胚肺成纤维细胞和单核细胞共同培养MMP-2活化酶和MMP-9活性和表达明显增强。 结论 肺癌H460细胞、胚肺成纤维细胞和单核细胞相互作用能通过上调MMP-2、MMP-9的表达促进肺癌的侵袭和转移。     

膜型基质金属蛋白酶-1对乳腺癌细胞浸润能力的影响

目的 研究膜型基质金属蛋白酶-1（MT1-MMP）对乳腺癌细胞株浸润能力的影响,并初步探讨其作用机制。方法 用20μg／ml刀豆素（ConA）刺激乳腺癌细胞株MDA—MB-453,促使其表达MT1-MMP蛋白,并用免疫细胞化学和Western blot法检测;然后加入外源性MMP-2原酶（proMMP-2）,并用明胶酶谱分析法检测proMMP-2被激活的情况;最后用侵袭实验检测细胞株的浸润能力。实验中将细胞株分为4组：空白对照组、ConA组、MMP-2组和ConA＋MMP-2组,各组实验结果进行对比分析。结果 利用ConA刺激后,ConA组和ConA＋MMP-2组的细胞均表达MT1-MMP蛋白,另两组无MT1-MMP蛋白表达。明胶酶谱分析实验发现,MMP-2组只检测到72000原酶形式的MMP-2,ConA＋MMP．2组同时检测到72000原酶形式和64000活酶形式的MMP-2,其余两组检测不到任何形式MMP-2。侵袭实验结果显示,ConA＋MMP-2组细胞穿过Marigel胶的细胞数目明显多于其他组。结论 MT1-MMP能显著增强乳腺癌细胞株的浸润能力,其机制主要是通过激活MMP-2原酶,降解肿瘤周围的基质成分实现的。     

Matrilysin over-expression in MCF-7 cells enhances cellular invasiveness and pro-gelatinase activation

Matrilysin (MMP-7) is over-expressed in various cancers and is thought to play important roles in tumor invasion and metastasis. However, the function of MMP-7 in breast cancer remains unclear. We therefore examined the expression of the MMP-7 gene in breast cancer (MCF-7) cells and the effect of its over-expression on cellular invasion. We transfected human MMP-7 into MCF-7 cells and selected clones that stably over-expressed the MMP-7 gene. The in vitro invasiveness of MCF-7 cells was quantified by use of the Matrigel invasion assay. Expression of MMP-7 mRNA was analyzed by quantitative RT-PCR. MMP secretion and activation were detected by gelatin zymography. We found that MMP-7-expressing clones had significantly increased invasion (P < 0.001), with increased MMP-7 expression and gelatinase activation as compared to the vector controls. We conclude that MMP-7 over-expression correlates with breast cancer in vitro invasiveness and that MMP-7 may promote invasion by increasing the secretion and activation of proMMP-2 and proMMP-9.     

Differential expression and activity status of MMP-1, MMP-2 and MMP-9 in tumor and stromal cells of squamous cell carcinomas of the lung.

Matrix metalloproteinases (MMPs) play a key role in cancer progression. Interstitial collagenase (MMP-1) and type IV collagenases (MMP-2, MMP-9) are involved in the initial breakdown of collagen and basement membrane components during tumor growth and invasion. Besides tumor cells, fibroblasts are especially involved in MMP production. The aim of this study was to quantify MMP-1, MMP-2 and MMP-9 within tumor cells and tumor-surrounding fibroblasts compared to normal lung epithelial cells to gain an insight into the function of these MMPs in squamous cell carcinomas of the lung. The expression and activity of MMP-1, MMP-2 and MMP-9 were analyzed in 30 squamous cell carcinomas and in normal lung tissue from the same patients by immunohistology and gelatin zymography. The majority of tumor cells were positive for MMP-1 (mean +/- SD: 67.3 +/- 26.7%) and MMP-9 (64.7 +/- 22.8%), whereas a significantly lower percentage of normal bronchoepithelial cells (47.3 +/- 25.4 and 40.3 +/- 24.2%, respectively; p < 0.01) and fibroblasts located in the tumor-surrounding tissue (39.7 +/- 14.3 and 38.1 +/- 24.1%, respectively; p < 0.01) expressed these MMPs. Only a few tumor cells showed any immunoreactivity for MMP-2 (4.4 +/- 6.7%), whereas a higher percentage of fibroblasts tested positive for this enzyme (8.6 +/- 13.1%; p < 0.01). Using gelatin zymography, we could demonstrate that MMP-2 is activated in the tumor only, not in normal lung tissue. The coordinated expression of MMP-1, MMP-2 and MMP-9 in tumor cells and/or their induction in tumor-surrounding fibroblasts and further activation in the tumor tissue may be involved in the high invasive and metastatic potential of squamous cell carcinomas of the lung. Comparing the results from immunohistology and zymography can give indications for distribution and activity of proteinases, especially certain MMPs such as MMP-2.     

PTEN磷酸酶活性对乳腺癌细胞ZR-75-1迁移能力的影响

目的:探求PTEN蛋白的磷酸酶活性对乳腺癌细胞ZR-75-1转移能力的影响。方法:采用脂质体介导法分别将野生型PTEN质粒(wt-PTEN)、磷酸酶失活的PTEN质粒(G129R-PTEN)和只具有蛋白磷酸酶活性的PTEN质粒(G129E-PTEN)转染PTEN基因缺失的人乳腺癌细胞株ZR-75-1,Western印迹法检测PTEN蛋白及P397-FAK的表达水平,体外细胞划痕实验观察PTEN磷酸酶活性对ZR-75-1细胞迁移能力的影响,细胞基质黏附试验和人工重组基底膜侵袭试验测定PTEN质粒转染和未转染的ZR-75-1细胞的黏附抑制率和侵袭抑制率,免疫组化法检测MMP-2的水平。结果:wt-PTEN、G129R-PTEN及G129E-PTEN3种质粒均成功转染ZR-75-1细胞并有PTEN蛋白的表达,其中wt-PTEN、G129E-PTEN均能抑制ZR-75-1细胞迁移;wt-PTEN和G129E-PTEN转染细胞之间的黏附抑制率和侵袭抑制率或侵袭细胞相对数均无显著性差异,但与G129R-PTEN转染的和未经转染的ZR-75-1细胞相比有显著性差异(P<0.01)。wt-PTEN和G129E-PTEN质粒转染的ZR-75-1细胞其P397-FAK水平均显著低于G129R-PTEN质粒转染的ZR-75-1细胞;wt-PTEN与G129E-PTEN质粒转染的ZR-75-1细胞MMP-2水平对比于G129R-PTEN质粒转染的和未经质粒转染的ZR-75-1细胞有显著性差异(P<0.01)。结论:具有双特异磷酸酶活性的野生型PTEN基因和只具蛋白磷酸酶活性的PTEN基因均能抑制乳腺癌细胞ZR-75-1的迁移,而磷酸酶失活的PTEN基因则无此作用。     

乳腺癌MMP-2、MMP-9的体外阻断研究

目的:探讨乳腺癌MMP-2、MMP-9表达的临床病理联系及直接阻断MMP-2、MMP-9对乳腺癌细胞增殖、侵袭及细胞周期的影响。方法:应用免疫组织化学方法检测48例乳腺癌MMP-2、MMP-9的表达。明胶酶谱法检测乳腺癌MCF-7细胞MMP-2、MMP-9的表达。MCF-7细胞接种于预铺BME的Tran-swell小室上室,与MMP-2、MMP-9的阻断剂CTT共孵育16h,观察阻断MMP-2、MMP-9对MCF-7细胞侵袭能力的影响。MCF-7细胞与CTT共孵育12h后,MTT法检测阻断MMP-2、MMP-9对细胞增殖能力的影响。MCF-7细胞与CTT共孵育24h后,PI染色流式法检测阻断MMP-2、MMP-9对细胞凋亡和细胞周期的影响。结果:48例乳腺癌组织有MMP-2表达者45例(93.75%),MMP-9表达者38例(79.17%),MMP-2和MMP-9在发生淋巴结转移组明显高表达(P<0.05);组织学分级越高,MMP-2和MMP-9的表达越显著(P<0.05)。明胶酶谱法检测:在MCF-7培养上清中检测到MMP-2、MMP-9的表达。在浓度为100μg/ml和200μg/ml时CTT对MCF-7细胞的侵袭抑制率达到39.5%和61.9%。在终浓度为50、100、200μg/ml时,CTT对MCF-7细胞的增殖率均无明显下降(P>0.05)。CTT处理组MCF-7细胞经流式法检测未见凋亡峰,细胞周期各期未见明显阻滞(P>0.05)。结论:MMP-2、MMP-9与乳腺癌的淋巴结转移密切相关,阻断MMP-2、MMP-9可有效抑制乳腺癌的浸润和转移。     

Suppression of Human Breast Cancer Cell Metastasis by Coptisine in Vitro

Background: Coptisine, an isoquinoline alkaloid extracted from Coptidis rhizoma, has many biologicalactivities such as antidiabetic, antimicrobial and antiviral actions. However, whether coptisine exerts anti-cancermetastasis effects remains unknown. Materials and Methods: Effects of coptisine on highly metastatic humanbreast cancer cell MDA-MB-231 proliferation were evaluated by trypan blue assay and on cell adhesion, migrationand invasion by gelatin adhesion, wound-healing and matrigel invasion chamber assays, respectively. Expressionof two matrix metalloproteinases (MMPs), MMP-9, MMP-2 and their specific inhibitors tissue inhibitor ofmetalloproteinase 1 (TIMP-1) and tissue inhibitor of metalloproteinase 2 (TIMP-2) were analyzed by RT-PCR.Results: Coptisine obviously inhibited adhesion to an ECM-coated substrate, wound healing migration, andinvasion through the matrigel in MDA-MB-231 breast cancer cells. RT-PCR revealed that coptisine reduced theexpression of the ECM degradation-associated gene MMP-9 at the mRNA level, and the expression of TIMP-1was up-regulated in MDA-MB-231 cells, while the expression of MMP-2 and its specific inhibitor TIMP-2 wasnot affected. Conclusions: Taken together, our data showed that coptisine suppressed adhesion, migration andinvasion of MDA-MB-231 breast cancer cells in vitro, the down-regulation of MMP-9 in combination with theincrease of TIMP-1 possibly contributing to the anti-metastatic function. Coptisine might be a potential drugcandidate for breast cancer therapy.     

七氟烷对乳腺癌BT549细胞侵袭、迁移及MMP-2、MMP-9表达的影响

目的:探讨七氟烷对乳腺癌BT549细胞侵袭、迁移及基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）表达的影响。方法:乳腺癌BT549细胞用七氟烷处理后,细胞划痕实验检测细胞迁移,Transwell小室检测细胞侵袭,Western blot检测MMP-2、MMP-9蛋白表达。结果:七氟烷处理后的BT549细胞迁移率由（36.45±3.21）%降低至（14.68±1.52）%,侵袭细胞数目由（136.58±13.36）个减少到（79.95±14.25）个,MMP-2水平由0.48±0.04减少为0.14±0.03,MMP-9水平由0.96±0.07减少为0.22±0.05,两组比较差异均具有统计学意义（P<0.05）。结论:七氟烷能够抑制乳腺癌BT549细胞迁移和侵袭,降低细胞中MMP-2、MMP-9表达水平。     

Dehydroepiandrosterone inhibits events related with the metastatic process in breast tumor cell lines

Dehydroepiandrosterone (DHEA), an adrenal hormone, has a protective role against cancer. We previously shown that DHEA inhibits the proliferation and migration of cell lines derived from breast cancer; however, the role of DHEA in others events related with these effects are unknown. We hypothesized that DHEA inhibits the expression of proteins and some events related with cell migration and metastasis. We determined the migration in Boyden chambers, the invasion in matrigel, anchorage-independent growth and the formation of spheroids in 3 cell lines (MCF-7, MDA-MB-231, ZR-75-30) derived from breast cancer exposed to DHEA. The secretion of metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), and several pro-inflammatory molecules in the secretome of these cells was also evaluated. DHEA inhibited the migration in transwells and the invasion in matrigel of MCF-7 and MDA-MB-231 cells. Besides, DHEA inhibited the anchorage-independent growth on agar and decreased the size of spheroids, and also reduced the secretion of IL-1α, IL-6, IL-8, and TNF-α in all cell lines. Metalloproteinase-1 (MMP-1) secretion was slightly decreased by DHEA treatment in MDA-MB-231 cells. Our results also showed that inhibition of migration and invasion induced by DHEA in breast cancer cells is correlated with the decrease of cytokine/chemokine secretion and the diminution of tumor cells growth. MCF-7 cells were the most responsive to the exposure to DHEA, whereas ZR-75-30 cells responded less to this hormone, suggesting that DHEA could be used in the treatment of breast cancer in early stages.     

miR-148a对肝癌细胞株侵袭和迁移的抑制作用及机制

背景与目的：原发性肝癌是肝细胞或肝内胆管上皮发生的恶性肿瘤,其复发和转移十分常见。本实验以慢病毒介导miR-148a,感染人肝癌SMMC-7721细胞,观察其对SMMC-7721细胞侵袭和迁移能力的影响。方法：选用SMMC-7721肝癌细胞株,进行miR-148a慢病毒载体的构建,筛选稳定表达miR-148a肝癌细胞株。RTPCR检测细胞中miR-148a的表达水平。划痕实验及Transwell侵袭实验检测细胞侵袭迁移能力。采用明胶酶谱法测定MMP-2、MMP-9的活性。蛋白质印迹法(Western blot)检测MMP-2、MMP-9和EMT相关蛋白(E-cadherin、vimentin)的表达情况。结果：RT-PCR结果显示,和对照组相比,LV-miR-148a肝癌细胞感染组表达miR-148a明显增高,体外侵袭实验细胞划痕实验显示过表达miR-148a后SMMC-7721的侵袭和迁移能力明显下降。明胶酶谱法结果显示过表达miR-148a的肝癌细胞,MMP-2和MMP-9降解明胶的能力下降(P<0.05)。过表达miR-148a同时能降低肝癌细胞SMMC-7721中vimentin、MMP-2和MMP-9蛋白的表达,但并未影响E-cadherin的表达。结论：miR-148a在体外能抑制肝癌细胞SMMC-7721的侵袭和迁移,其机制可能与下调MMP-2、MMP-9和vimentin的表达水平有关。     

塔斯品碱衍生物TPD7对乳腺癌细胞迁移和侵袭的抑制作用

目的:探讨塔斯品碱衍生物TPD7对乳腺癌细胞MCF-7和ZR-75-30细胞迁移和侵袭的影响,阐明TPD7抑制细胞迁移和侵袭可能的作用机制。方法:采用细胞划痕法、Transwell小室侵袭法检测MCF-7和ZR-75-30细胞的迁移和侵袭。采用Western blot法检测MMP9、β-catenin及c-Myc的蛋白表达。RT-PCR法检测β-catenin及c-Myc的mRNA表达。结果:TPD7对MCF-7细胞和ZR-75-30细胞迁移和侵袭均有显著抑制作用,且下调MMP9、β-catenin、c-Myc的蛋白表达和mRNA表达,呈剂量依赖性。结论:TPD7抑制乳腺癌MCF-7和ZR-75-30细胞迁移和侵袭,可能是通过Wnt信号通路抑制上皮-间质转化。     

miR-126下调MMP-2抑制人脑胶质瘤细胞侵袭

目的初步探讨miR-126抑制人胶质瘤细胞侵袭的可能机制。方法化学合成miR-126,脂质体转染人脑胶质瘤U87细胞,应用RT-PCR、Western blot检测MMP-2基因和蛋白的表达情况,并应用Transwell小室检测转染前后细胞侵袭力的变化。结果miR-126上调后U87细胞的MMP-2基因和蛋白表达降低,并且细胞侵袭力明显降低。结论化学合成的miR-126在抑制人胶质瘤细胞侵袭过程中发挥重要作用,可能成为胶质瘤基因治疗的新靶点。