醋酸甲地孕酮联合三氧化二砷对肝癌HepG2细胞株体外抑制作用的研究

目的 探讨醋酸甲地孕酮（MA）联合三氧化二砷（As2O3）对肝癌HepG2细胞株体外增殖的影响及其可能机制。方法 将对数生长期HepG2细胞分为4组：MA单药组、As2O3单药组、MA+As2O3联合组和对照组,根据前期研究结果确定药物剂量分别为75.0 μmol/L MA、1.0 μg/ml As2O3,对照组加入等量细胞培养液。24 h后采用CCK-8方法、流式细胞学检测及Western blotting检测MA单药组、As2O3单药组及MA+As2O3联合组对HepG2细胞增殖、细胞凋亡及X连锁凋亡抑制蛋白（XIAP）表达的影响。结果CCK-8法显示MA+As2O3组较MA或As2O3单药组均能够明显抑制HepG2细胞生长;流式细胞仪结果显示MA+As2O3联合组的细胞凋亡率高于MA或As2O3单药组。MA+As2O3作用后XIAP表达较MA或As2O3单药组降低。结论MA联合As2O3能够发挥对HepG2细胞协同抑制作用,其中诱导细胞凋亡可能是其机制之一。     

Ciglitazone对肝癌细胞株HepG2在体内外生长的影响

目的研究过氧化物酶体增殖物激活受体γ(peroxisome proliferator activated receptor γ,PPARγ)激动剂ciglitazone对人肝癌细胞株 HepG2体内、外生长的影响,并探讨其可能机制。方法培养HepG2细胞,加入不同浓度的ciglitazone,用生长曲线检测HepG2细胞体外生长情况,用流式细胞仪进行细胞周期分析; Western blot检测ciglitazone对PPARγ蛋白表达的影响。建立裸鼠肝癌动物模型,随机将其分为两组:对照组(A组,10只),ciglita zone注射组(B组,10只),B组隔天注射ciglitazone 100μ(100 μmol/L)连续15次,对照组注射100 μL生理盐水,1个月后,切除瘤灶、称瘤重、计算抑瘤率。标本用Western blot检测细胞周期素D1(cyelinD1)、细胞周期素依赖性激酶抑制子P21蛋白的表达。结果(1)ciglitazone体外抑制HepG2细胞生长,并呈现明显时间和剂量依赖性。(2)经ciglitazone处理后PPARγ蛋白表达增加。(3) 经ciglitazone治疗后,A组、B组的瘤重分别为3．73±0．22g,2．60±0．35g,B组的抑瘤率达30％,A组的cyclinD1较B组明显增高,A 组的P21蛋白表达水平较B组明显降低。结论 Ciglitazone能抑制肝癌HepG2细胞的恶性增殖,其抑制作用呈明显的时间及剂量依赖性,并诱导其分化,其机制与PPARγ干预细胞周期的调控有关。     

The growth inhibitory effect of mesenchymal stem cells on tumor cells in vitro and in vivo

Mesenchymal stem cells (MSCs) play an important role in the development and growth of tumor cells. The purpose of this study is to confirm the effect of MSCs on tumor cell growth in vitro and in vivo and to elucidate the mechanism. MSCs were isolated from mouse bone marrow and cocultured with murine hepatoma H22, lymphoma (YAC-1 and EL-4) and rat insulinoma INS-1 cell lines. The growth inhibitory effect of MSCs on tumor cells was tested through MTT and 3H-TdR incorporation assay. The apoptosis induction effect of MSCs on tumor cells was assessed with flow cytometry (FCM) and RT-PCR assay. MSCs were inoculated into BALB/c mice alone or coinoculated with ascitogenous hepatoma cells intraperitonealy, respectively. The tumor growth inhibition of MSCs was investigaed through the incidence and volume of ascites formation, and the immunosuppression effect was studied with splenocyte response to ConA stimulation test and T cell subsets analysis (FCM). The results showed that MSCs exhibited a number-dependent growth inhibitory effect on murine tumor cell lines in vitro and inhibited the growth of ascitogenous hepatoma cells in vivo without host immunosuppression. MSCs could upregulate tumor cells mRNA expression of cell cycle negative regulator p21 and apoptosis associated protease caspase 3. The findings of this experimental study demonstrated that MSCs had potential inhibitory effects on tumor cell growth in vitro and in vivo without host immunosuppression, by inducing apoptotic cell death and G(0)/G(1) phase arrest of cancer cells.     

Effect of megestrol acetate or melengestrol acetate on preneoplastic and neoplastic mammary growth in mice

Pellets of megestrol acetate and melengestrol acetate were implanted subcutaneously every one and two months, respectively, into 3-5 month old SHN virgin mice. Spontaneous mammary tumourigenesis was significantly enhanced by both steroids. The formation of preneoplastic mammary hyperplastic alveolar nodules (HAN) was, in contrast, inhibited by both progestins. As these results closely resemble the effects of progesterone and of medroxyprogesterone acetate on the development of mammary tumours both in the SHN mouse and in the woman, it seems likely that this animal model may prove valuable for the study of human breast cancer that is no longer responsive to progestins.     

Effects of megestrol acetate on growth and secretion of a pituitary tumor

The effect of the administration of the6-methylated progestagen megestrol acetate on the growth of the estrogen-induced transplantable PRL/ACTH-secreting rat pituitary tumor7315a was investigated. A significant inhibition of tumor growth was observed only during the last week of a four week period of drug administration (6 mg/kg). Administration of megestrol acetate suppressed the total content and the ability to release PRL, ACTH and LH of the pituitary gland of tumor bearing rats. Further, a significant suppression of the anterior pituitary, ovarian and uterine weights was observed in the megestrol acetate treated animals. Megestrol acetate did not exert a direct effect on PRL, ACTH and LH secretion by normal rat pituitary glands incubated in vitro.We conclude that megestrol acetate exerts in the model used a suppressive effect on PRL, ACTH and LH release and led to a diminution of the pituitary content of these hormones, while it has also anti-estrogenic and anti-androgenic properties. The effects of the drug help further explain its beneficial effects in metastatic breast, endometrial, renal and prostatic cancer in man.     

The effect of GM-CSF and G-CSF on the growth of human osteosarcoma cells in vitro and in vivo

The human osteosarcoma cell line, MG63, responds both to GM-CSF and to G-CSF in vitro. To assess the significance of these observations to tumor growth in vivo, MG63 cells were engineered by retroviral infection to produce human GM-CSF or G-CSF. These retrovirally infected cells become autostimulatory as measured by increased [³H]-thymidine incorporation (3-to 7-fold) and anchorage-independent colony formation (7-to 10-fold) as compared with uninfected MG63 cells or cells infected with control (neo^r) retrovirus. The increased proliferation induced by exogenous GM-CSF or G-CSF on uninfected MG63 cells in both assays could be completely inhibited by anti-GM-CSF or anti-G-CSF antibodies, while the same antibodies only partially abrogated proliferation by the growth-factor-producing cells. None of 34 nude or SCID mice developed tumors when injected s.c. with uninfected or neo^r-virus-infected cells. In contrast, all 30 mice injected with GM-CSF-or G-CSF-producing MG63 cells developed tumors which were G418-resistant and factor-producing. Tumor cell DNA showed a polyclonal retroviral integration pattern indistinguishable from that in the DNA of cells injected into mice. Tumors that formed following injection of a mixture of G418-resistant, GM-CSF-producing cells and cells infected with virus containing only the hygro^r gene contained hygromycin-resistant cells in the same proportion as was present in the original cell mixture. These data indicate that GM-CSF and G-CSF can support the growth of an osteosarcoma cell line both in vitro and in vivo whether the factor is supplied by autocrine production or from exogenous sources.     

Oncogene transfection of mink lung cells: effect on growth characteristics in vitro and in vivo

Three sublines have been derived from the parental line Mv1Lu by transfection with normal and mutated Ha-ras, and myc oncogenes, and subsequent cloning. All the oncogenes have increased the growth rate of the cell in vitro, increased their plating efficiency in monolayer and suspension, and reduced their serum dependence. Growth in vivo as xenografts in nude mice has also been increased. Very few tumours were generated from the parental line and those that did form did so after a prolonged lag period, while the transfected lines produced tumours with 100% efficiency, and a short lag period. In general the effects of ras transfection were more extreme, with the highest growth rates and plating efficiencies in vitro and the shortest lag period and doubling times in vivo. There was no increase in plasminogen activator activity as a result of transfection, and the invasive behaviour of the lines in organotypic culture was broadly similar.     

CIK细胞联合贝伐单抗对肝癌HepG2细胞的体内外抗瘤活性

目的:探讨细胞因子诱导杀伤(CIK)细胞联合贝伐单抗(bevacizuma)对肝癌HepG2细胞的体内外抗肿瘤活性及其作用机制.方法:提取健康供血者外周血的单个核细胞(PBMC),加入多种细胞因子促进CIK细胞成熟,在流式细胞仪上进行CIK细胞的免疫表型分析.CIK细胞与贝伐单抗单独或联合作用于HepG2细胞后,利用CCK-8测定其对HepG2细胞体外增殖活性的影响;利用侵袭小室(Transwell)和划痕实验测定其对HepG2细胞侵袭迁移活性的影响;Western blotting检测HepG2细胞Akt和Erk信号通路相关蛋白磷酸化的变化.建立HepG2细胞裸鼠皮下移植瘤模型,并随机分为生理盐水组、CIK组、贝伐单抗组及CIK细胞联合贝伐单抗组,给药28 d后处死裸鼠,剥取瘤体,免疫组化法检测瘤组织CD31及Ki67蛋白的表达.结果:提取健康人PBMC诱导14 d后,CIK细胞表型分析显示CD3+ CD56+细胞扩增达(36.33±2.58)％.与单独治疗组比较,联合组对HepG2细胞的抗肿瘤增殖活性显著增强(P ＜0.05);CIK细胞和贝伐单抗两药联合比单独给药组对HepG2细胞的侵袭[(75.6 ±9.53) vs (304.8 ±45.73)、(359.8 ±38.10)个,P＜0.01]和迁移[(29.35±8.14)％vs (55.07±6.27)％、(60.50±9.73)％,P＜0.05]能力的抑制更强;CIK细胞和贝伐单抗两药单独及联合都能抑制Akt和Erk的磷酸化;CIK联合贝伐单抗组可显著抑制移植瘤生长以及移植瘤组织平均血管密度和Ki67表达,与单独治疗及对照组细胞比较差异有统计学意义(P＜0.05).结论:CIK联合贝伐单抗在体内外对HepG2细胞增殖、侵袭、迁移均有抑制作用,且优于单独治疗组.     

Radiosensitizing effect of medroxyprogesterone acetate on endometrial cancer cells in vitro

From clinical experience it is known that medroxyprogesterone acetate (MPA) can increase the radiosensitivity of adenocarcinomas of the corpus uteri. This study investigates this phenomenon in vitro. Primary explants of highly differentiated adenocarcinomas were irradiated with or without pretreatment with MPA and compared with an untreated control group and to a group treated with MPA only. Cell culture itself was performed on an agarose medium in order to prevent overgrowth by fibroblasts. Untreated samples formed 43 +/- 5 clones, explants treated with MPA only produced 39 +/- 5 clones, a difference which was not statistically different; samples irradiated without pretreatment produced 16 +/- 8 and samples after combined treatment 9 +/- 3 clones (all values means +/- SD). This numeric reduction of cell growth through preirradiation treatment with MPA was statistically significant. The effect of MPA as a radiosensitizer may be due to its potential to prolong the radiosensitive G2 phase of the cell cycle. This effect of MPA may be useful also in other hormone-dependent tumors.     

The effect of beta-elemene on alpha-tubulin polymerization in human hepatoma HepG2 cells

Objective： To investigate the impact of beta-elemene injection on the growth and alpha-tubule of human hepatocarcinoma HepG2 cells. Methods： Cell proliferation was assessed by MTT assay. Cell cycle distribution was detected by flow cytometry （FCM）. The mRNA expression of alpha-tubulin was measured by RT-PCR. Western blot analysis was used to determine protein expression of alpha-tubulin and the polymerization of tubulin. Results： Beta-elemene injection inhibited HepG2 cells proliferation in a dose- and time-dependent manner; FCM analysis indicated beta-elemene injection induced cell cycle arrested at S phase. RT-PCR and western- blot analysis showed that beta-elemene injection down-regulated alpha-tublin at both mRNA and protein levels, presenting a dose-dependent manner. Moreover, beta-elemene injection reduced the polymerization of microtubules in a dose-dependent manner. Conclusions： Beta-elemene injection can inhibit the proliferation of hepatoma HepG2 cells and induce cell apoptosis, the mechanism might be partly related to the down-regulation of alpha-tubulin and inhibition of microtubular polymerization.     

Brivanib 对 HepG2细胞的抑制作用及机制

目的：探讨 Brivanib 对人 HepG2肝癌细胞的增殖抑制作用及其机制。方法：MTS 法检测 Brivanib 对HepG2细胞的增殖抑制作用,流式细胞仪检测不同浓度 Brivanib 处理后 HepG2细胞的凋亡率,Western blot 检测凋亡相关蛋白 Bcl -2、Bax 的表达情况,免疫荧光观察 Brivanib 处理后 HepG2细胞内源性 LC3表达情况, Western blot 检测自噬关键蛋白 LC - I 向 LC - II 的转换及 p62的表达。结果：与空白对照组相比,Brivanib 对HepG2肝癌细胞的增殖抑制率随剂量增加和时间延长而增加,呈剂量和时间依赖性。不同浓度 Brivanib 作用72h 后 HepG2细胞的凋亡率分别为（13.06±4.06）%、（20.89±1.83）%、（44.29±2.56）%,其诱导凋亡与下调 Bax、上调 Bcl -2表达相关。2.5μmol/ L Brivanib 处理 HepG2细胞48h 后内源性 LC3表达增加。Western blot 分析表明,LC3- I 向 LC3- II 转换增加,p62表达降低。结论：Brivanib 抑制 HepG2人肝癌细胞增殖,诱导凋亡和自噬活化。     

Inhibitory effect of S-adenosylmethionine on the growth of human gastric cancer cells in vivo and in vitro

Background and Objective: S-adenosylmethionine (SAM), the most important methyl donor in human body, is generally used to treat cholestasis in clinic. In recent years, SAM has been found to have inhibitory effects on breast cancer, liver cancer and colon carcinoma. This study was to investigate the inhibitory effects of SAM on human gastric cancer cells in vivo and in vitro, and the antitumor mechanisms. Methods: The effects of SAM on the proliferation of gastric cancer SGC-7901 and MKN-45 cells were determ...     

TMPyP4对肝癌细胞HepG2生长抑制作用的研究

目的 TMPyP4是G-四链体的配体,能够诱导形成G-四链体结构并增强G-四链体结构的稳定性,加强G-四链体作为负调控因子抑制MET基因的表达;G-四链体结构的形成可以选择性的抑制肿瘤细胞增殖.本研究旨在分析TMPyP4对肝癌细胞HepG2生长抑制作用的研究,为MET高表达的肿瘤靶向治疗提供新思路.方法 采用CCK 8方法分析HepG2细胞增殖变化,通过Hoechst 33342细胞核染色和Annexin V-FITC/PI双染流式细胞术分析HepG2细胞凋亡水平,以及利用流式细胞术分析HepG2细胞周期分布的变化.结果 CCK-8分析HepG2细胞增殖显示,与对照组比较,10 μmol/L TMPyP4对HepG2细胞存活率无显著影响,20、50、100和200 μmol/L TMPyP4组HepG2细胞存活率分别为(82.28±1.99)％、(68.88±3.06)％、(55.57±2.76)％和(50.32±5.78)％,F=120.542,P＜0.001,说明高于20 μmol/L的TMPyP4显著抑制细胞增殖;流式细胞术分析细胞凋亡不明显,只有高浓度TMPyP4诱导少量HepG2细胞发生了凋亡,并出现了坏死细胞;100和200 μmol/L TMPyP4的实验组细胞经Hoechst 33342染色,有少数HepG2细胞表现出细胞核固缩、致密,形态不规则,染色不均一,呈现凋亡表现;不同浓度的TMPyP4处理HepG2细胞使G0/G1期细胞增加了20％,S期细胞减少,G2/M期细胞也减少,表示HepG2细胞发生了G0/G1细胞周期阻滞.结论 G-四链体的配体TMPyP4显著抑制了肝癌细胞HepG2的细胞增殖,诱导少量HepG2细胞凋亡,并使HepG2细胞发生了G0/Gl细胞周期阻滞.     

甲地孕酮对晚期恶性肿瘤患者生活质量改善的观察

目的：观察甲地孕酮对晚期恶性肿瘤患者生活质量改善的作用。方法：120例晚期肿瘤患者随机分为A、B两组，A组60例，采用甲地孕酮分散片（megestrolacetate，MA，宜利治）与化疗联用；B组60例，采用单独化疗。结果：生活质量改善A组优于B组；疼痛疗效观察：A组总有效率为71．7％与B组相比有显著性差异；不良反应：III、IV度毒副反应出现率A组少于B组，A、B两组有效率（CR＋PR）分别为38．3％、35．O％，两组间的比较无显著性差异。疾病控制率（CR＋PR＋SD）A组（76．7％）比B组（58．3％）明显提高。中位生存期（MST）A组较B组长48天（P〈0．05）。结论：甲地孕酮联合化疗治疗晚期恶性肿瘤副作用小，有较大的临床应用价值，可作为晚期恶性肿瘤治疗期间改善生存质量的有效手段。     

Bisphosphonates inhibit the growth of mesothelioma cells in vitro and in vivo.

PURPOSE: Bisphosphonates (such as risedronate and zoledronate) are widely used inhibitors of bone resorption. Despite their in vitro antiproliferative effects in various cancer cells, bisphosphonates have not exhibited significant antitumor efficacy in animal models of visceral cancer, which may be due to their poor bioavailability. The diagnostic use of radioactive bisphosphonates has revealed the accumulation of bisphosphonates in mesothelioma, which prompted us to test the antitumor efficacy of bisphosphonates in this disease. EXPERIMENTAL DESIGN AND RESULTS: Treatment with either risedronate or zoledronate (2 x 10(-4) to 2 x 10(-6) mol/L) inhibited the growth of AB12 and AC29 mouse mesothelioma cells and induced the accumulation of unprenylated Rap1A in these cells. Both these in vitro effects were reversed by geranygeraniol, an end product of the mevalonate pathway that these bisphosphonates inhibit. Both bisphosphonates also induced the phosphorylation of the p38 mitogen-activated protein kinase in AB12 and AC29 cells. The inhibition of p38 augmented bisphosphonate-induced growth inhibition in these cells. Bisphosphonate-induced p38 phosphorylation was not reversible by geranylgeraniol. Risedronate (15 mg/kg) and zoledronate (0.5 mg/kg) inhibited the growth of s.c. tumors and increased the median survival of mice with i.p. mesothelioma tumors in vivo. Discussion: In conclusion, risedronate and zoledronate inhibit the mevalonate pathway and induce p38 activation in mesothelioma cells in vitro. The effects on the mevalonate pathway dominate because the net result is growth inhibition. Both bisphosphonates also inhibit mesothelioma tumor growth in vivo and prolong the survival of mesothelioma-bearing mice. These results support further study of bisphosphonates in the management of mesothelioma.     

RNAi-mediated ERK2 knockdown inhibits growth of tumor cells in vitro and in vivo

The MAPK MEK/ERK pathway is often upregulated in cancer cells and represents an attractive target for development of anticancer drugs. Only few data concerning the specific functions of ERK1 and 2 are reported in the literature. In this report, we investigated the specific role of ERK1 and 2 in liver tumor growth both in vitro and in vivo. DNA synthesis and cells in S phase analysed by flow cytometry, correlated with strong inhibition of Cdk1 and cyclin E levels, are strongly reduced after exposure to the MEK inhibitor, U0126. We obtained a significant reduction of colony formation in soft agar assays and a reduction in the size of tumor xenografts in nude mice treated with U0126. Then, we could specifically abolished ERK1 or 2 expression by small-interfering RNA (siRNA) and demonstrated that ERK2 knockdown but not ERK1 interferes with the process of replication. Moreover, we found that colony formation and tumor growth in vivo were significantly inhibited by targeting ERK2 using stable chemically modified siRNA. Taken together, our results emphasize the importance of the MEK/ERK pathway in liver cancer cell growth in vitro and in vivo and argue for a crucial role of ERK2 in this regulation.     

葫芦素B在体内外对乳腺癌细胞的生长抑制作用

目的:体内外观察葫芦素B(cucurbitacin B)对人乳腺癌细胞系MCF-7的增殖抑制和诱导凋亡作用,为临床应用提供依据.方法:用浓度分别为0.001、0.01、0.1、1、10以及100μmoL/L的葫芦素B处理MCF-7细胞,24、48和72 h后用MTT法检测细胞增殖.用O.1和10μmol/L的葫芦素B处理MCF-7细胞24h,之后用流式细胞仪检测细胞周期分布和细胞凋亡率,并用倒置显微镜和荧光显微镜观察细胞凋亡.建立乳腺癌裸鼠移植瘤模型,观察葫芦素B的体内抑瘤作用.结果:MTT结果显示葫芦素B对MCF-7细胞的增殖抑制作用具有明显的剂量和时间依赖性.流式细胞仪检测提示,随着葫芦素B浓度增高,处于S期和G2/M期细胞的比例逐渐升高,同时伴随G0/G1期细胞的减少以及细胞凋亡率的逐渐升高.统计学分析提示,各实验组之间以及实验组与对照组之间均有显著差异(P相似文献   

二甲基亚砜(DMSO)对人肝癌HepG2 细胞的诱导分化作用的实验研究

目的:探讨二甲基亚砜(DMSO)对人肝癌HepG2细胞的诱导分化作用。方法:体外培养人肝癌HepG2细胞,用DMSO干预细胞,MTT法检测药物对细胞增殖活力的影响;倒置相差显微镜和瑞氏-姬姆萨染色观察细胞形态学的变化;放射免疫法检测细胞甲胎蛋白（AFP）和清蛋白(ALB)的分泌量;酶促反应试剂盒检测细胞中γ-谷胺酰转肽酶（γ-GT）、碱性磷酸酶（ALP）的活性。结果:用DMSO处理HepG2细胞72h后,能显著抑制细胞的增殖（P＜0.05）;细胞的形态向成熟细胞分化;细胞AFP的分泌量和γ-GT酶的活性明显降低,ALP活性和ALB含量则显著升高（P＜0.05）。结论:DMSO具有诱导人肝癌HepG2向正常细胞分化的作用并抑制细胞增殖。