三氧化二砷对H22肝癌荷瘤小鼠免疫功能及瘤体的影响

目的 探讨三氧化二砷(As2O3)对H22肝癌荷瘤小鼠免疫功能的影响及实体瘤生长情况.方法 利用流式细胞仪及改进的MTT还原法分别对正常小鼠、荷瘤小鼠对照组、荷瘤小鼠As2O3治疗组进行T细胞亚群及NK细胞活性(NKCA)的测定,并比较各组瘤重,计算瘤重抑制率.结果 肝癌小鼠荷瘤对照组与正常组比较,CD3+,CD4+,CD4+/CD8+及NKCA均明显减少(P<0.01);低剂量及高剂量 As2O3治疗组与荷瘤对照组比较,CD3+, CD4+, CD4+/CD8+及NKCA均明显升高(P<0.05),CD8+无明显变化;而高剂量 As2O3治疗组与低剂量 As2O3治疗组比较,CD4+,NKCA明显升高(P<0.05),CD3+, CD4+/CD8+无明显变化.治疗组瘤重抑制率分别为39.12%,45.74%,病理观察可见间质内大量炎症细胞浸润.结论 AS2O3 能抑制实体肿瘤的生长,提高肝癌小鼠的免疫功能,这是一值得继续研究的临床课题.     

三氧化二砷治疗原位移植肝癌的药效学研究

目的:研究三氧化二砷(As2O3)对原位移植小鼠肝癌抑制作用及免疫功能的影响.并探讨As2O3对小鼠肿瘤细胞H22的抑制机理.方法:建立小鼠H22肝原位移植模型,考察As2O3对小鼠原位肝癌抑制率的影响,对免疫器官指数一脾指数(SI)、胸腺指数(TI)的影响,流式细胞术检测对细胞周期和免疫因子的影响,HE染色进行病理学评价,并作综合评价.结果:As2O3高剂量和低剂量的抑瘤率分别为44.60%和26.98%;流式细胞术检测可见As2O3使小鼠肝癌细胞阻滞在S期,从而影响癌细胞的增殖,同时As2O3可调节荷瘤动物IL-2、TNF-a的水平,而对IL-12无明显影响.结论:As2O3能够抑制小鼠原位移植肝癌的生长,抑制移植小鼠肿瘤细胞H22的增殖,改善荷瘤动物的免疫功能.     

消癌解毒方抑制肝癌H22移植瘤的生长及其机制

目的:研究消癌解毒方(Xiaoai Jiedu Recipe,XJR)对肝癌H22细胞小鼠移植瘤的抑制作用及其可能的机制。方法:建立小鼠H22移植瘤模型,分对照组和XJR低、中、高剂量(10、30、90g/kg)组及顺铂(0.001g/kg)组进行治疗,检测各组移植瘤生长情况,H-E染色观察各组移植瘤组织的病理改变。流式细胞术检测各组移植瘤细胞周期及凋亡率,ELISA法测定各组移植瘤小鼠外周血VEGF水平。结果:与对照组比较,XJR各剂量以及顺铂对H22移植瘤的生长均具有显著的抑制作用,抑瘤率分别为24.5%、42.8%、21.1%、58.6%(P<0.05或P<0.01)。病理观察见中剂量XJP组和顺铂组移植瘤组织大片状坏死,有明显染色质固缩环。中剂量XJR和顺铂将移植瘤组织细胞阻滞于G0/G1期(P<0.05),S期细胞数明显减少(P<0.05)。中剂量XJR组移植瘤细胞的凋亡率与顺铂组相当[(60.52±6.40)%vs(71.32±16.02)%,P>0.05]。中、高剂量XJR组和顺铂组小鼠外周血VEGF水平均显著降低[(104.3±6.1)、(105.8±7.2)、(88.6±4.3)vs(120.7±12.6)ng/ml,P<0.05或P<0.01]。结论:XJR能抑制H22小鼠移植瘤的生长,促进移植瘤细胞凋亡、抑制VEGF的产生可能是其抗癌作用机制之一。     

三氧化二砷-泊洛沙姆４０７缓释剂瘤内注射治疗裸鼠人肝癌移植瘤的实验研究

目的：观察三氧化二砷-泊洛沙姆４０７（Ａｓ_２Ｏ_３-Ｐ４０７）缓释剂对裸鼠皮下人肝癌移植瘤的有效性。方法：以人肝癌ＨｅｐＧ２细胞为来源建立裸鼠皮下移植瘤模型,实验共分３组：Ａｓ_２Ｏ_３-Ｐ４０７缓释剂治疗组、Ａｓ_２Ｏ_３治疗组和生理盐水（ＮＳ）对照组,于造模成功后６～８天行瘤体注射,每４天１次,共４次。比较各组抗癌疗效,观察裸鼠体重、肿瘤体积变化、抑瘤率、肿瘤组织病理及骨髓涂片以及裸鼠的存活情况、生存质量。结果：Ａｓ２Ｏ３ Ｐ４０７缓释剂治疗组肿瘤生长受到显著抑制,肿瘤体积较Ａｓ_２Ｏ_３组小（Ｐ＜０.０５）,瘤重分别为（０.２５７±０.０８０）ｇ和（０.６４１±０.１０９）ｇ,差异有统计学意义（Ｐ＜０.０５）;两组与ＮＳ对照组瘤重（１.０９４±０.１４７）ｇ相比,差异有统计学意义（Ｐ＜０.０５）。治疗后Ａｓ_２Ｏ_３-Ｐ４０７缓释剂组裸鼠体重大于Ａｓ_２Ｏ_３组和ＮＳ对照组,差异有统计学意义（Ｐ＜０.０５）。各组均未见骨髓抑制现象。结论：Ａｓ_２Ｏ_３-Ｐ４０７缓释剂瘤内注射,使用安全,可维持局部有效药物浓度、延长作用时间,疗效优于Ａｓ_２Ｏ_３注射液。     

三氧化二砷联合热疗抑制大鼠肝癌的体内实验研究

目的：研究热疗联合三氧化二砷（As2O3）对大鼠肝癌多药耐药性的影响,探讨其作为肝癌辅助治疗的可行性。方法：采用含有walker-256肿瘤细胞的大鼠腹水稀释后接种于大鼠腹股沟皮下,7天后取下瘤块并切成（1．0—2．0）mm3大小的小块,然后将小瘤块接种于50只大鼠的左肝叶上,10天后将大鼠分成4组,分别为对照组,热疗组,As2O3组及（热疗＋As2O3）组,同时给予治疗,治疗结束后第2天处死大鼠。测量瘤重分析As2O3联合热疗的体内抑瘤作用;用H—E染色法观察肿瘤细胞病理象变化;免疫组化s—P法检测肿瘤组织内Pgp（P—glycoprotein,P-糖蛋白）的表达情况。结果：热疗组,As2O3组及（热疗＋As2O3）组的平均瘤重均低于对照组（P〈0．05）;同时（热疗＋As2O3）组与As2O3组及热疗组相比较,前者抑瘤率高于后两者（P〈0．05）。在实验过程中,对照组荷瘤鼠活动减少,毛发稀疏,进食水少,而热疗组大鼠活动稍差,其余两组基本正常,饮食状况无明显改变,体重未见明显减轻。各组细胞均有坏死,大部分为凝固性坏死。对照组肿瘤细胞生长比较旺盛,坏死面积较小。其他三组坏死都比较彻底,并且呈现大片状坏死,尤以（热疗＋As2O3）组明显,镜下呈一片荒凉景象,肿瘤细胞溶解,破碎,多见细胞核碎裂及核固缩表象。免疫组化法观察到多药耐药细胞呈片状分布,Pgp主要分布于耐药细胞的胞膜面,少数分布于胞浆当中;Pgp显色指数在对照组与各处理组之间差异显著（P〈0．05）,且（热疗＋As2O3）组较其他处理组差异显著（P〈0．05）。结论：As2O3和热疗在体内对大鼠肝癌均有明显的抑制作用,两者联合应用有协同作用,其作用机制可能与抑制多药耐药基因（MDR1）的表达有关。     

三氧化二砷联合热疗抑制大鼠肝癌的体内实验研究

目的:研究热疗联合三氧化二砷(As2O3)对大鼠肝癌多药耐药性的影响,探讨其作为肝癌辅助治疗的可行性。方法:采用含有walker-256肿瘤细胞的大鼠腹水稀释后接种于大鼠腹股沟皮下,7天后取下瘤块并切成(1.0-2.0)mm3大小的小块,然后将小瘤块接种于50只大鼠的左肝叶上,10天后将大鼠分成4组,分别为对照组,热疗组,As2O3组及(热疗+As2O3)组,同时给予治疗,治疗结束后第2天处死大鼠。测量瘤重分析As2O3联合热疗的体内抑瘤作用;用H-E染色法观察肿瘤细胞病理象变化;免疫组化S-P法检测肿瘤组织内Pgp(P-glycoprotein,P-糖蛋白)的表达情况。结果:热疗组,As2O3组及(热疗+As2O3)组的平均瘤重均低于对照组(P<0.05);同时(热疗+As2O3)组与As2O3组及热疗组相比较,前者抑瘤率高于后两者(P<0.05)。在实验过程中,对照组荷瘤鼠活动减少,毛发稀疏,进食水少,而热疗组大鼠活动稍差,其余两组基本正常,饮食状况无明显改变,体重未见明显减轻。各组细胞均有坏死,大部分为凝固性坏死。对照组肿瘤细胞生长比较旺盛,坏死面积较小。其他三组坏死都比较彻底,并且呈现大片状坏死,尤以(热疗+As2O3)组明显,镜下呈一片荒凉景象,肿瘤细胞溶解,破碎,多见细胞核碎裂及核固缩表象。免疫组化法观察到多药耐药细胞呈片状分布,Pgp主要分布于耐药细胞的胞膜面,少数分布于胞浆当中;Pgp显色指数在对照组与各处理组之间差异显著(P<0.05),且(热疗+As2O3)组较其他处理组差异显著(P<0.05)。结论:As2O3和热疗在体内对大鼠肝癌均有明显的抑制作用,两者联合应用有协同作用,其作用机制可能与抑制多药耐药基因(MDR1)的表达有关。     

肿瘤抑素抑制肝癌移植瘤生长的实验研究

目的 观察肿瘤抑素慢病毒载体对裸鼠肝癌移植瘤生长的抑制作用,探讨其在肝癌基因治疗中的应用前景。方法 建立人肝癌细胞株SMCC-7721裸鼠皮下移植瘤模型,当肿瘤直径达到0.3～0.5cm时分别在瘤周及瘤体内多点注射100μl磷酸盐缓冲液（PBS组）、5×109 TU慢病毒绿色荧光蛋白（Lenti-GFP组）和5×109 TU 肿瘤抑素重组慢病毒（Lenti-Tum组）。观察3组的肿瘤体积变化,采用RT-PCR和Western blotting法分别检测3组肿瘤组织中肿瘤抑素的mRNA和蛋白水平,并采用Kaplan-Meier法进行生存分析。结果 注射21天后,Lenti-Tum组的肿瘤体积为（702.1±143.7）mm3,小于PBS组的（1622.2±253.8）mm3和Lenti-GFP组的（1538.5±284.9）mm3（P＜0.05）;Lenti-Tum组的抑瘤率为56.7％,显著高于Lenti-GFP组的5.2％（P＜0.05）;Lenti-Tum组中肿瘤抑素的mRNA和蛋白水平均高于其他两组。Lenti-Tum组小鼠的中位生存时间为100天,明显高于PBS组的58天和Lenti-GFP组的59天（P＜0.05）。结论 肿瘤抑素经慢病毒转染能够明显抑制肝癌移植瘤的生长,延长荷瘤裸鼠的生存时间。     

小鼠肝癌固化瘤苗的免疫实验研究

目的 探讨肝癌瘤苗对小鼠免疫功能的影响。方法 测定小鼠抑瘤率及胸腺细胞功能变化。结果 用瘤苗免疫后小鼠能抗同源10~7个癌细胞攻击,其中40％全部有效,60％部分有效。结论 此瘤苗有明显抑瘤作用,且此作用与其影响胸腺细胞代谢增强有密切关系。     

三氧化二砷抗肝癌作用的研究进展

 临床已证实三氧化二砷（As2O3）能有效治疗急性早幼粒细胞性白血病，在此基础上，有关As2O3抗肝癌作用的研究报告也越来越多，该文就As2O3抗肝癌作用的实验研究和临床应用作一综述。     

Inhibition by arsenic trioxide of human hepatoma cell growth

Arsenic trioxide (As(2)O(3)) has been shown to be effective for treatment of patients with refractory or relapsed acute promyelocytic leukemia and a variety of other malignant hematopoetic disorders. We studied the effect of this agent on proliferation of human hepatoma-derived cell lines (SK-Hep-1, HepG2, and HuH7). In HuH7 cells, As(2)O(3) reduced proliferation time- and dose-dependently at 1 and 2 microM, while in SK-Hep-1 and HepG2 cells, As(2)O(3) inhibited proliferation at 2 and 4 microM respectively. Cell cycle analysis by flow cytometry showed that As(2)O(3) induced apoptosis in these hepatoma-derived cells as confirmed by appearance of sub-G(1) cells. Sensitivity of hepatoma-derived cells to As(2)O(3) was inversely related to their intracellular glutathione (GSH) and intensity of GSH synthesis. Arsenic sensitivity was restored to relatively resistant cell lines when GSH was depleted by L-buthionine sulfoximine (BSO). These results indicate that As(2)O(3) may have therapeutic potential for treatment of hepatocellular carcinoma.     

三氧化二砷诱导肝癌细胞凋亡的初步研究

目的：研究三氧化二砷（Ａｓ２Ｏ３）对肝癌细胞株Ｂｅｌ－７４０２的抑制增殖效应及凋亡诱导作用。方法：采用ＭＴＴ法检测药物对细胞的抑制作用、相差显微镜和电镜观察细胞形态变化、琼脂糖凝胶电泳测定ＤＮＡ Ｌａｄｄｅｒ、流式细胞术检测细胞周期变化。结果：各浓度Ａｓ２Ｏ３组（０．２５～１６μｍｏｌ／Ｌ）均可抑制肝癌细胞增殖,且抑制率具有浓度和时间依赖性,同作用时间呈直线相关,其中浓度组为１０μｍｏｌ／Ｌ）均可抑制肝癌细胞增殖,且抑制率具有浓度和时间依赖性,同作用时间呈直线相关,其中浓度组为１０μｍｏｌ／Ｌ抑制率最大,９６ｈ时达８３．８９％,２μｍｏｌ／Ｌ Ａｓ２Ｏ３以抑制肝癌细胞的增殖为主,但在形态及生化方面未见凋亡改变;１０μｍｏｌ／Ｌ Ａｓ２Ｏ３作用１０ｈ可见形态学变化,４８ｈ可见ＤＮＡ Ｌａｄｄｅｒ出现。流式细胞检查     

PTEN在三氧化二砷诱导肝癌细胞(SMMC-7721)凋亡中的作用

目的 研究PTEN在三氧化二砷(arsenic trioxide,As2O3)诱导肝癌细胞凋亡中的作用.方法 采用四甲基偶氮唑盐(MTT)方法检测As2O3对SMMC-7721细胞增殖的抑制作用;流式细胞仪定量检测各组细胞的凋亡率;Western blot方法检测各组细胞中PTEN蛋白的表达情况;用PTEN siRNA转染SMMC-7721细胞,观察其对As2O3诱导细胞凋亡作用的影响.结果 As2O3能显著抑制SMMC-7721细胞存活率,其抑制率呈剂量依赖关系;As2O3剂量依赖性诱导肝癌细胞凋亡;Western blot检测显示As2O3明显增加细胞中PTEN蛋白的表达;PTEN siRNA转染可减弱As2O3对SMMC-7721细胞凋亡的诱导作用.结论 As2O3诱导SMMC-7721细胞凋亡可能与其诱导PTEN表达有关.     

三氧化二砷对肝癌细胞survivin基因和端粒酶活性的影响

目的:探讨三氧化二砷(As2O3)诱导肝癌细胞凋亡时survivin基因表达和端粒酶活性的变化。方法:采用倒置相差显微镜观察细胞形态,流式细胞仪分析细胞亚二倍体百分率,逆转录-多聚酶链式反应检测细胞survivin mRNA表达,免疫组织化学染色检测细胞survivin蛋白的表达,端粒酶检测试剂盒检测端粒酶活性。结果:0.25~2.00μmol/L As2O3明显抑制肝癌Bel-7402细胞生长,诱导细胞凋亡,survivin mRNA和蛋白表达上调,端粒酶活性下降,呈时间、剂量依赖关系。结论:As2O3抑制肝癌Bel-7402细胞增殖,诱导细胞凋亡,可能与其降低端粒酶活性有关;而survivin基因表达上调,可能是抵抗As2O3诱导凋亡的途径之一。     

Berberine enhances inhibition of glioma tumor cell migration and invasiveness mediated by arsenic trioxide

Background  

Arsenic trioxide (As₂O₃) exhibits promising anticarcinogenic activity in acute promyelocytic leukemic patients and induces apoptosis in various tumor cells in vitro. Here, we investigated the effect of the natural alkaloid berberine on As₂O₃-mediated inhibition of cancer cell migration using rat and human glioma cell lines.     

抑制端粒酶活性对As2O3诱导肝癌细胞凋亡的影响

目的： 〖HT5"SS〗观察端粒酶反义寡核苷酸、As2O3对肝癌细胞生长的影响,寻找低剂量、低毒、高效、安全的抗肝癌新疗法。〖HT5W〗方法：〖HT5"SS〗 自行设计合成靶向端粒酶模板区的20 nt硫代反义寡核苷酸,观察端粒酶反义寡核苷酸与As2O3对肝癌细胞端粒酶活性的影响及两者协同对肝癌细胞生长的影响,采用HE染色、流式细胞仪及DNA琼脂糖凝胶电泳观察端粒酶反义寡核苷酸与As2O3对肝癌细胞的凋亡诱导作用,以流式细胞术检测肝癌细胞的Fas、FasL、bcl2蛋白的表达。〖HT5W〗结果： 〖HT5"SS〗5 μmol/L的端粒酶反义寡核苷酸作用24h可显著抑制肝癌细胞端粒酶活性（P<0.01）;肝癌细胞端粒酶活性被抑制后对As2O3诱导凋亡的敏感性显著增加,表现为低浓度、短时间即可诱导肝癌细胞凋亡（P<0.01）,端粒酶反义寡核苷酸与As2O3对肝癌细胞的凋亡诱导作用是通过Fas、FasL途径实现的。〖HT5W〗结论：〖HT5"SS〗抑制肝癌细胞端粒酶活性可显著增强其对As2O3诱导凋亡的敏感性,减少砷剂用量,两者协同有潜在的临床应用前景。     

Opposing effects of arsenic trioxide on hepatocellular carcinomas in mice

Arsenic trioxide (As2O3) is a potent antitumor agent used to treat acute promyelocytic leukemia (APL) and, more recently, solid tumors. However, the dose of As2O3 required to suppress human xenographs in mice is markedly higher than that used to treat APL in humans. Paradoxically, low doses of As2O3 stimulate angiogenesis, which might be expected to promote tumor growth. Clearly, appropriate dosages of As2O3 are required to treat human patients to avoid toxicity and undesirable side effects. In the present study, we investigated As2O3 with respect to its toxicity and effects on tumor growth, angiogenesis and cell apoptosis using H22 hepatocellular carcinoma (HCC) cells in a mouse model of HCC. As2O3 inhibited tumor growth and angiogenesis, and enhanced tumor cell apoptosis at doses greater than 1 mg/kg, but mice lost weight and failed to thrive at doses of 4 mg/kg and greater. In contrast, low doses (<1 mg/kg) of As2O3 promoted tumor growth, upregulated the expression of vascular endothelial growth factor and tumor angiogenesis, and had no effect on tumor cell apoptosis. In vitro studies demonstrated that As2O3 inhibited the proliferation of H22 tumor cells and bovine aortic endothelial cells, and induced their apoptosis in a dose- and time-dependent fashion, suggesting that the mechanism of As2O3-mediated inhibition of tumor growth is due to direct effects of the drug on both tumor cells and endothelia. In summary, different doses of As2O3 have opposing effects on tumor growth and angiogenesis. The results demonstrate that As2O3 has a narrow window of therapeutic opportunity with respect to dosage, and that low doses of the drug as used in metronomic therapy should be used with extreme caution.     

Induction of apoptosis and inhibition of human gastric cancer MGC-803 cell growth by arsenic trioxide

Arsenic trioxide (As2O3), used to treat human diseases for centuries in traditional Chinese medicine, has been identified as a very effective antileukaemic agent, but its effect on solid tumours which could be more suitable for clinical treatment with arsenic compounds is still unknown. In this study, we investigated the in vitro effect of As2O3 at concentrations of 0.01-1 microM against six human malignant cell lines, MGC-803, HIC, MCF-7, HeLa, BEL-7402 and A549 cells. As2O3 inhibited growth and induced apoptosis in these malignant cells at varying degrees, in a time dose-dependent manner. The most marked effects were seen in the gastric cancer cell line, MGC-803. In contrast, minimal growth inhibition and induction of apoptosis occurred in human embryonic pulmonary cells following treatment with As2O3 found at the same concentrations. Changes in intracellular Ca2+, following As2O3 treatment were measured by Ca2+ sensitive fluorescent probe Indo-1/AM in flow cytometric assays. The increase in intracellular Ca2+ correlated with the sensitivity of these cells to As2O3, possibly indicating that a critical intracellular Ca2+ signal transduction pathway could be involved in As2O3-mediated cell-death and its selectivity. The marked sensitivity of MGC-803 cells in vitro suggests that As2O3 may be a potential antigastric cancer agent.     

Galectin-3 inhibition sensitizes human renal cell carcinoma cells to arsenic trioxide treatment

The anti-tumor effects of arsenic trioxide (ATO) were well established in acute promyelocytic leukemia, but not in renal cell carcinoma (RCC). Recent evidences indicate that galectin-3 (Gal-3) plays an anti-apoptotic role in chemotherapy induced tumor cell death. This study was intended to clarify the exact roles of Gal-3 performed in ATO-induced apoptosis in RCC cells. Weak apoptosis was observed in Gal-3-positive RCC cells (Caki-1, Caki-2, 786-0, and ACHN) following ATO treatment. However, ATO treatment upregulated Gal-3 expression concurrently caused a Synexin-cooperated translocation of Gal-3 from the nucleus to the cytoplasm. Gal-3-knockdown cells were more sensitive to ATO treatment as indicated by a strong mitochondria-dependent apoptosis following ATO treatment. Meanwhile, Gal-3 was found to inhibit ATO-induced apoptosis through enhancing Bcl-2 expression and stabilizing mitochondria. To confirm the results obtained from genetic method, we employed a Gal-3 inhibitor, modified citrus prectin (MCP), and co-treated the RCC cells with ATO. The cells showed an increased apoptosis in the syngeneic application of Gal-3 inhibition and ATO compared with ATO application alone. Based on these results, we conclude that Gal-3 inhibition sensitizes human renal cell carcinoma cells to ATO treatment through increasing mitochondria-dependent apoptosis. Our studies implicate synergetic application of ATO and Gal-3 inhibition as a potential strategy for RCC treatment.     

三氧化二砷治疗真性红细胞增多症的初步研究

目的：在观察Ａｓ２Ｏ３对Ｋ５６２细胞系和真性红细胞增多症（ＰＶ）患者外周血单个核细胞的体外效应的基础上,分析Ａｓ２Ｏ３治疗３例ＰＶ患者的临床结果。方法：以Ｋ５６２细胞和ＰＶ患者的新鲜细胞为体外靶子,通过细胞形态学,流式细胞仪和Ａｎｎｅｘｉｎ－Ｖ的测定检测细胞凋亡。每日静脉滴注１０ｍｌ０．１％Ａｓ２Ｏ３注射液治疗３例ＰＶ患者,至完全缓解停药并继续随访。结果：１￣２μｍｏｌ／Ｌ Ａｓ２Ｏ３注射液诱导Ｋ