18β-GA对人高转移卵巢癌HO-8910PM细胞增殖、黏附和侵袭能力的影响

目的观察18β-甘草次酸（18β-GA）对人高转移卵巢癌HO-8910PM细胞增殖、黏附和侵袭能力及黏附斑激酶（FAK）、基质金属蛋白酶（MMP）-9表达的影响。方法将18β-GA作用于HO-8910PM细胞,采用MTT法检测HO-8910PM细胞增殖抑制率;细胞黏附试验检测细胞黏附能力;Transwell chamber法检测细胞侵袭能力;Westernblot法检测细胞FAK、MMP-9蛋白表达水平。结果 18β-GA明显抑制高转移卵巢癌细胞HO-8910PM的生长增殖、黏附和侵袭能力;Western blot法检测表明药物能下调FAK、MMP-9蛋白的表达。结论 18β-GA可抑制HO-8910PM细胞黏附、侵袭,其机制可能是通过下调FAK、MMP-9蛋白表达而进行的。     

急性髓性白血病细胞对内皮细胞表达细胞间黏附分子-1的影响

急性髓细胞性白血病（AML）患者可因白细胞过高或白细胞淤滞导致多脏器衰竭，与白血病细胞黏附有关。研究显示白血病细胞表面黏附分子表达异常。目前认为细胞间黏附分子（ICAM-1）在正常白细胞外渗过程中起主要作用，IL-1β，TNFα和IFM等细胞因子直接介导内皮细胞ICAM-1的合成。而在AML中，影响内皮细胞ICAM-1表达因素的体外实验研究较少。因此，我们观察AML白血病细胞培养上清对内皮细胞表达ICAM-1的作用，旨在研究AML白血病细胞对内皮细胞的影响。     

人卵巢癌细胞HABP1的表达及其与侵袭转移能力的相关性

目的探讨透明质酸结合蛋白1（HABP1）在卵巢癌侵袭转移过程中的作用。方法分别用荧光实时定量RT—PCR和Western blot法检测HABP1 mRNA和HABP1蛋白在人卵巢癌细胞株HO-8910及高转移HO-8910PM细胞株中的表达,采用体外侵袭实验测定细胞侵袭能力。结果HO-8910PM中HABP1 mRNA和蛋白的表达水平均高于HO-8910,HO-8910PM的侵袭转移能力高于HO-8910。结论HABP1的高表达可能在人卵巢癌细胞的侵袭转移过程中发挥一定作用。     

GHGKHKNK八肽对肿瘤细胞克隆形成、黏附及体外侵袭能力的影响

目的探讨GHGKHKNK八肽对小鼠黑色素瘤B16-F10细胞克隆形成与体外侵袭能力的作用及其机制。方法用不同实验方法观察GHGKHKNK八肽对克隆形成能力、细胞侵袭能力及GHGKHKNK八肽对层黏连蛋白受体（LN-R）,细胞黏附分子-1（ICAM-1）,内皮组织钙黏连素（E-cadherin）在小鼠黑色素瘤B16-F10细胞中表达情况。结果不同剂量GHGKHKNK八肽对小鼠黑色素瘤细胞B16-F10的克隆形成侵袭均有抑制作用,与对照组相比较均有显著性差异;小鼠黑色素瘤细胞B16-F10经免疫细胞化学染色后,可见小鼠黑色素瘤细胞B16-F10LN-R、ICAM-1、E-cadhe-sin表达降低。结论GHGKHKNK八肽可以明显抑制肿瘤细胞的克隆形成、黏附和侵袭能力。     

黏着斑激酶在结肠癌细胞中的表达及对其迁移的影响

[目的]观察局部黏着斑激酶（FAK）在结肠癌细胞中的表达及对结肠癌细胞侵袭动力和生长的影响。[方法]应用RT-PCR、Western blot方法测定结肠癌细胞株（HT-29）和正常肠上皮细胞中FAK的表达；在血小板源性生长因子（PDGF）刺激肿瘤细胞后，应用细胞黏附分析和体外侵袭试验方法，测定不同时间点细胞的体外黏附能力变化及侵袭能力，同时行FAK表达测定。[结果]结肠癌细胞较正常上皮细胞存在较高水平的FAK表达。PDGF干预组与非干预组比较，能促进HT-29的黏附功能（90．0％）和侵袭能力（54．5％），在该过程中，FAK的表达和活性升高（P〈0．05）。[结论]FAK对结肠癌细胞的体外黏附与迁移发挥重要的作用。     

麝黄消瘤方抑制肝癌细胞侵袭粘附作用的实验研究

目的：观察麝黄消瘤方（SHF）兔含药血清对SMMC-7721肝癌细胞侵袭粘附能力的影响及其机制的探讨。方法：以体外培养的肝癌细胞株为研究对象，将肝癌细胞株分别接种于中药组、对照组血清中培养，绘制细胞生长曲线，计算细胞生长抑制率。以流式细胞仪（FCM）测SHF对肝癌细胞表面细胞间粘附分子-1（ICAM-1）表达的影响。以免疫组化法观察SHF对肝癌细胞抑癌基因nm23-H1蛋白水平的表达。以细胞侵袭实验观察SHF对与纤维连接蛋白（FN）粘附的SMMC-7721细胞侵袭运动能力及其生长状况。以岍法测量SHF对肝癌细胞与FN粘附的影响。结果：中药组血清较对照组明显抑制肝癌细胞的体外生长，在第5天抑制率最高达66．25％；中药处理的肝癌细胞ICAM-1表达明显低于对照组，nm23-H1蛋白水平明显高于对照组。SHF可抑制肝癌细胞的侵袭运动能力及其与FN之间的粘附。结论：SHF可抑制SMMC-7721细胞的侵袭粘附能力，其作用与中药血清上调nm23-H1蛋白水平和降低ICAM-1表达等有关。     

熊果酸对卵巢癌细胞黏附、运动和侵袭的影响

目的探讨熊果酸(UA)对人高转移卵巢癌细胞HO-8910黏附、运动和侵袭的影响。方法以不同浓度的UA处理高转移卵巢癌细胞HO-8910,采用MTT法及细胞黏附人工重组基底膜实验检测UA对HO-8910细胞的细胞毒作用及黏附能力的影响;Transwell小室法检测UA对HO-8910细胞侵袭能力和趋化运动能力的影响。结果不同浓度UA处理后,随着UA浓度从10μmol/L增加到30μmol/L,与对照组比较,对细胞黏附抑制率增强(P0.01),能显著降低HO-8910运动能力和侵袭能力(P0.01)。结论 UA能抑制卵巢癌细胞HO-8910黏附、运动和侵袭能力。     

不同亚型胃癌细胞株裸鼠移植瘤中组织蛋白酶B的表达

目的：研究组织蛋白酶B（CB）在裸鼠移植瘤中的表达及其与不同亚型胃癌细胞株体内成瘤性的关系。方法：以体外层粘素（laminin)附粘法筛选获得的两株MKN45胃癌细胞亚株：对层粘素高粘附力（Lm^ )的细胞亚株和低粘附力（Lm^-)的细胞亚株建立人胃癌裸鼠移植瘤模型，应用免疫组化及原位杂交方法观察荷瘤组织中CB的表达情况。结果：Lm^ 细胞株瘤块与周围组织粘连较Lm^-瘤块明显。免疫组化和原位杂交均显示移植瘤组织中的CB表达增强，且Lm^ 瘤块的表达强于Lm^-瘤块，以肿瘤边缘表达最强。结论：CB在胃癌细胞株移植瘤中的表达增强，尤以Lm^ 细胞株为著，其表达上调可能与癌细胞株的体内侵袭行为有关。     

细胞间黏附分子与胃癌侵袭、转移关系的研究进展

细胞间黏附分子-1(intercellularadhesionmolecule-1,ICAM-1)是体内重要的细胞活性分子,不仅参与了机体免疫过程及炎症反应,而且还通过与其相应配体的结合,介导癌细胞与不同细胞、基质的黏附,最终使癌细胞逃避免疫监视,利于侵袭转移.近年来,对ICAM-1与恶性肿瘤侵袭转移关系的研究已成为研究热点,其在肿瘤侵袭转移中的作用日益受到众多学者的重视.现就其近年的文献从ICAM-1的结构、生物学特征、在胃癌组织中的表达及作用机制等几个方面作一综述.     

过表达MiR-381调控LRH-1-MMP-2通路抑制卵巢癌细胞转移的机制

目的探讨人卵巢癌组织中miR-381的表达及对卵巢癌细胞迁移和侵袭的影响。方法聚合酶链反应(PCR)检测卵巢癌组织中miR-381表达水平;慢病毒转染卵巢癌SKVO3细胞构建miR-381稳定过表达细胞株;通过划痕愈合试验、Transwell侵袭小室检测细胞迁移及侵袭情况,通过裸鼠尾静脉注射肿瘤细胞建立肿瘤肺转移模型,检测过表达miR-381对肿瘤细胞体内转移的影响;PCR及Western印迹法检测miR-381潜在靶点肝受体同源物(LRH)-1的表达变化。结果 miR-381在卵巢癌组织中表达水平明显低于癌旁组织(P0.05);上调miR-381的表达能够抑制卵巢癌SKVO3细胞体外迁移(P0.05)及侵袭能力(P0.05);在体内,上调miR-381的表达对SKVO3细胞裸鼠肺转移瘤的形成具有抑制效应(P0.05);机制上,过表达miR-381后显著抑制LRH-1在SKVO3细胞内的表达水平(P0.05),并抑制了基质金属蛋白酶(MMP)-2表达水平(P0.05)。结论 miR-381在卵巢细胞癌中表达降低,上调miR-381能够通过抑制LRH-1的表达进而抑制MMP-2表达而发挥抗肿瘤转移作用。     

KAI1 gene suppresses invasion and metastasis of hepatocellular carcinoma MHCC97‐H cells in vitro and in animal models

Background: Downregulation of KAI1 gene expression has been found in many types of cancer cells and is closely related to cancer invasion and metastasis. This study was aimed at investigating the effects and possible underlying mechanisms of KAI1 gene on invasion and metastasis of human hepatocellular carcinoma (HCC). Methods: The invasive ability, visco‐elastic properties and cell adhesion forces were analysed in different HCC cells originating from the MHCC97‐H cell line transfected with either the sense or the antisense KAI1 expression plasmid. Tumuorigenicity, metastatic abilities, extracellular matrix (ECM) and intercellular adhesion molecule‐1 (ICAM‐1) expression were also evaluated in the nude mouse models of the xenografted and orthotopic liver cancer cells. Results: Compared with their parental cells, in the HCC cells transfected with the sense KAI1 gene, the invasive ability in vitro was significantly decreased (P<0.01); the cellular elastic coefficients K₁, K₂ and μ were significantly higher (P<0.05); the cells adhesion forces to fibronectin were significantly lower (P<0.01). The sense KAI1 gene transfection into the cancer cells also inhibited their invasion and lung metastasis in the orthotopic liver cancer nude mice. However, the opposite changes were observed in the HCC cells transfected with the antisense KAI1 gene. KAI1 gene transfection also affected ECM and ICAM‐1 expression in the transplanted liver cancer. Conclusion: The KAI1 gene plays an important role in the invasion and metastasis of human HCC and its upregulation in HCC cells suppresses their invasive and metastatic abilities. KAI1 gene functioned as a metastasis inhibitor by regulating the HCC cell biophysical behaviours including aggregation, adhesion, motility and visco‐elastic properties.     

Prognostic value of intercellular adhesion molecule (ICAM)-1 expression in breast cancer

Purpose  

The intercellular adhesion molecule (ICAM)-1 is expressed on many cell types including endothelial cells and different cancer cell entities. Experimental data strongly indicate that ICAM-1 can activate intracellular signalling pathways in cancer cells leading to enhanced cell motility, invasion and metastasis. Yet, little is known about the role of ICAM-1 expression during malignant progression in breast cancer patients.     

Higher CO2-insufflation pressure inhibits the expression of adhesion molecules and the invasion potential of colon cancer cells

AIM： To investigate the influence of CO2-insufflation pressure on adhesion, invasion and metastatic potential of colon cancer cells based on adhesion molecules expression. METHODS： With an/n vitro artificial pneumoperitoneum model, SW1116 human colon carcinoma cells were exposed to CO2-insufflation in 5 different pressure groups： 6 mmHg, 9 mmHg, 12 mmHg, 15 mmHg and control group, respectively for 1 h. Expression of E-cadherin, ICAM-I, CD44 and E-selectin was meas- ured at 0, 12, 24, 48 and 72 h after CO2-insufflation using flow cytometry. The adhesion and invasion capacity of SW1116 cells before and after exposure to CO2-insufflation was detected by cell adhesion/invasion assay in vitro. Each group of cells was injected intraperitoneally into 16 BALB/C mice. The number of visible abdominal cavity tumor nodules, visceral metas-tases and survival of the mice were recorded in each group. RESULTS： The expression of E-cadherin, ICAM-1, CD44 and E-selectin in SWl116 cells were changed significantly following exposure to CO2 insufflation at different pressures （P 〈 0.05）. The expression of E-cadherin, CD44 and ICAM-1 decreased with increasing CO2-insufflation pressure. The adhesive/ invasive cells also decreased gradually with increasing pressure as determined by the adhesion/invasion assay. In animal experiments, the number of abdominal cavity tumor nodules in the 15 mmHg group was also significantly lower than that in the 6 mmHg group （29.7± 9.91 vs 41.7±14.90, P = 0.046）. However, the survival in each group was not statistically different. CONCLUSION： CO2-insufflation induced a temporary change in the adhesion and invasion capacity of cancer cells in vitro. Higher CO2-insufflation pressure inhibited adhesion, invasion and metastatic potential in vitro and in vivo, which was associated with reduced expression of adhesion molecules.     

四种细胞黏附分子在老年宫颈癌浸润中的作用

目的探讨细胞黏附分子在老年宫颈癌局部浸润中的作用。方法应用组织芯片技术和免疫组织化学方法检测子宫颈鳞状细胞原位癌30例、微小浸润癌34例、浸润癌36例中PECAM-1、ICAM-1、E-cadherin、P-选择素的表达。结果PECAM-1在原位癌中的表达率明显低于微小浸润癌、浸润癌(P<0.05),微小浸润癌、浸润癌中表达率无明显差别;P选-择素的表达率在原位癌、微小浸润癌、浸润癌中无明显差别;ICAM-1、E-cadherin在原位癌中的阳性表达率均明显高于微小浸润癌中的表达率(P<0.05),微小浸润癌中的阳性表达率均明显高于浸润癌(P<0.05)。ICAM-1、E-cadherin在微小浸润癌、浸润癌中的表达水平之间存在密切相关(P<0.01),ICAM-1、E-cadherin的阳性表达与子宫颈癌的浸润呈明显负相关(r=-0.872 6,P<0.01和r=-0.796 8,P<0.01)。结论PECAM-1、ICAM-1、E-cadherin、P-选择素都参与了老年宫颈癌的局部浸润,其中ICAM-1、E-cadherin在局部浸润过程中发挥重要作用。     

Activation of p70S6K induces expression of matrix metalloproteinase 9 associated with hepatocyte growth factor-mediated invasion in human ovarian cancer cells

The expression of hepatocyte growth factor (HGF) receptor, encoded by the Met oncogene, is elevated in ovarian and a variety of cancers. Here we show that human ovarian cancer cells with high Met expression were more sensitive to the cell motility and invasion effect of HGF. Met down-regulation by small interfering RNAs or K252a resulted in reduced migration in response to HGF. The invasive/migratory phenotype activated by HGF can be blocked by specific inhibitors of the phosphatidylinositol-3-kinase (PI3K) cascade, inhibitor of p70(S6K), and also the expression of a dominant-negative Akt, demonstrating that HGF transmits the motogenic signal through PI3K and Akt to p70(S6K). A significant role for p70(S6K) in cell invasion is further supported by the observation that expression of constitutively active forms of p70(S6K) is sufficient to induce invasive and migratory phenotypes in ovarian cancer cells. Importantly, activation of p70(S6K) stimulated expression and proteolytic activity of matrix metalloproteinase (MMP)-9 and cellular invasion, whereas it had little effect on MMP-2, suggesting for the first time that MMP-9 up-regulation by p70(S6K) as a key step for HGF-induced invasion and migration. These data suggest that interfering p70(S6K) may provide a novel means of controlling tumor cell invasiveness.     

Steroid receptor coactivator-3, a homolog of Taiman that controls cell migration in the Drosophila ovary, regulates migration of human ovarian cancer cells

Border cell migration is a process that occurs during Drosophila ovarian development in which cells derived from a simple epithelium migrate and invade neighboring tissue. This process resembles the behavior of cancerous cells that derive from the simple epithelium of the human ovary. One important regulator of border cell migration is Taiman, a homolog of steroid receptor coactivator-3 (SRC-3). Because increasing evidence indicates that similarities exist between the molecular control of migration of border cells and of cancer cells, we investigated whether SRC-3 controls ovarian cancer cell migration. Little or no SRC-3 expression was detected in normal ovarian surface epithelium, ovarian cysts and borderline ovarian tumors that lack stromal invasion. In contrast, SRC-3 was abundantly expressed in high-grade ovarian carcinomas. Inhibiting SRC-3 expression in ovarian cancer cells markedly reduced cell spreading and migration, and altered intracellular localization of focal adhesion kinase. This inhibitory effect on cell migration was independent of the estrogen receptor (ER) status of the cells. These studies reveal a novel role for SRC-3 in ovarian cancer progression by promoting cell migration, independently of its role in estrogen receptor signaling.     

Genistein inhibits invasive potential of human hepatocellular carcinoma by altering cell cycle, apoptosis, and angiogenesis

AIM: To study the in vitro and in vivo inhibitory effects of genistein on invasive potential of Bel 7402 hepatocellular carcinoma (HCC) cells and to explore the underlying mechanism. METHODS: Bel 7402 HCC cells were exposed to genistein. The invasive activity of tumor cells was assayed in transwell cell culture chamber. p125FAK expression and cell cycle were evaluated by a functional assay. Cell apoptosis analysis was performed with TUNEL method. In addition, bilateral subrenal capsule xenograft transplantation of HCC was performed in 10 nude mice. Genistein was injected and the invasion of HCC into the renal parenchyma was observed. Microvessels with immunohistochemical staining were detected. RESULTS: Genistein significantly inhibited the growth of Bel 7402 cells, the inhibitory rate of tumor cells was 26 -42%. The invasive potential of Bel 7402 cells in vitro was significantly inhibited, the inhibitory rate was 11-28%. Genistein caused G2/M cell cycle arrest, S phase decreased significantly. The occurrence of apoptosis in genistein group increased significantly. The expression of p125FAK in 5 μg/mL genistein group (15.26±0.16%) and 10 μg/mL genistein group (12.89±0.36%) was significantly lower than that in the control group (19.75±1.12%,P<0.05). Tumor growth in genistein-treated nude mice was significantly retarded in comparison to control mice, the inhibitory rate of tumor growth was about 20%. Genistein also significantly inhibited the invasion of Bel 7402 cells into the renal parenchyma of nude mice with xenograft transplant. The positive unit value of microvessels in genistein-treated group (10.422±0.807) was significantly lower than that in control group (22.330±5.696, P< 0.01). CONCLUSION: Genistein can effectively inhibit the invasive potential of Bel 7402 HCC cells by altering cell cycle, apoptosis and angiogenesis, inhibition of focal adhesion kinase may play a significant role in this process.     

Expression of intracellular adhesion molecule-1 and vascular cell adhesion molecule-1 and homing factor CD44 after engraftment of Graves' lymphocytes in xenotransplanted human thyroid tissue in athymic nude mice.

The expression of adhesion molecules on thyrocytes and endothelium cells plays an important role in the pathogenesis of Graves' disease (GD). The intercellular adhesion molecule-1 (ICAM-1), the vascular cell adhesion molecule-1 (VCAM-1), and the homing receptor CD44 are responsible for the specific migration of lymphocytes in autoimmune thyroid diseases (AITD) (homing). Eight weeks after transplantation of thyroid tissue from 26 patients with nonautoimmune thyroid disease (nontoxic nodular goiter [NTG]) into nude mice, peripheral (PBL) and intrathyroidal lymphocytes (ITL) from 14 patients with NTG and 12 patients with GD were grafted into the animals. Two days after lymphocyte engraftment, the thyroid transplants were examined histologically (HE) and immunohistologically stained with monoclonal antibodies directed against ICAM-1, VCAM-1, and CD44. After injection of GD lymphocytes, thyroid transplants expressed significantly more ICAM-1, VCAM-1, and CD44 than after injection of NTG lymphocytes. This expression was even more pronounced after grafting of GD intrathyroidal lymphocytes. Our data demonstrate that only GD lymphocytes induce the expression of adhesion molecules and homing factor CD44, both of which play an important role in the migration of lymphocytes and induction of the autoimmune process.     

Integrin β3 down-regulates invasive features of ovarian cancer cells in SKOV3 cell subclones

Purpose  To investigate the role of integrin β3 in invasive features of ovarian cancer SKOV3 cells, by comparing different metastatic subclones. Methods  In the present study, two cell subclones, termed as S1 and S21, which possessed high and low metastatic potential, respectively, were isolated and established from human ovarian cancer parental cell line SKOV3 by the limited dilution method. The expressions of integrin αv, integrin αvβ3, integrin β3, E-cadherin, FAK and ILK in the two cell subclones were compared by means of real-time RT-PCR or flow cytometry. Subsequently, S21 was transfected with siRNA for integrin β3 and the effects of transfection were examined by methyl thiazolyl tetrazolium (MTT) assay, colony formation assay, Matrigel invasion assay and cell migration assay. Results  The expressions of integrin αvβ3, integrin β3 and E-cadherin were markedly down-regulated in S1; however, there were no significant differences in the expressions of integrin αv, FAK and ILK β. Of note, more than 70% knockdown of integrin β3 expression was obtained by siRNA technique. The integrin β3-siRNA-transfected cells showed significant increases in cell proliferation, cell migration and invasive activity in contrast with the mock-transfected cells. The expressions of integrin αvβ3 and E-cadherin were lower in the integrin β3-siRNA-transfected cells compared to the mock control. Conclusion  Integrin β3, like E-cadherin, may be also a suppressor gene down-regulating invasive features of ovarian cancer cells in SKOV3 cell subclones.