异种肿瘤细胞疫苗抗肿瘤活性的研究

目的研究异种肿瘤细胞疫苗抗肿瘤免疫反应,从而达到抑瘤作用.方法采用福氏佐剂与人肺腺癌AGZY-138、人肝癌HepG2细胞分别混合研磨制成疫苗,小鼠腹部皮下多点注射免疫小鼠或治疗小鼠,定期测定瘤体,计算抑瘤率、生存率等.结果预防保护实验肿瘤接种第五周时,与对照组比较抑瘤作用显著,肿瘤治疗实验肿瘤接种第五周时,人肺癌AGZY-138疫苗对小鼠Lewis肺癌治疗作用与对照组比较有显著意义,而人肝癌HepG2疫苗治疗作用对小鼠H22肝癌作用不显著,与对照组比较无差异.结论用异种肿瘤细胞疫苗对小鼠进行主动免疫后,再接种肿瘤,肿瘤生长明显减慢.肺癌疫苗预防保护小鼠生存期延长.     

逆癌酮抗小鼠肺癌作用的初步研究

背景与目的　研究发现丹参酮具有抗肿瘤作用,逆癌酮是以丹参酮为主要成分并加入其它抗肿 瘤成分的中药制剂。本研究拟探讨逆癌酮对小鼠Lewis肺癌的抑癌作用及其机制。方法　用逆癌酮进行体 内抑癌实验,并用流式细胞术检测肿瘤组织中细胞凋亡指数和细胞周期分布。结果　共进行两次实验,每次 实验单一性别小鼠。逆癌酮组与5 Fu组的瘤重与对照组比较均有显著性差异(P<0.05),逆癌酮对小鼠 Lewis肺癌的抑瘤率分别为38.9%和32.2%,5 Fu对小鼠Lewis肺癌的抑瘤率分别为59.6%和53.9%;各 组间肺部转移率均无显著性差异(P>0.05)。流式细胞仪分析表明,逆癌酮组细胞凋亡指数(11.22%)明显 高于对照组(5.43%)(P<0.05),而细胞周期分布与对照组比较无显著性差异(P>0.05)。结论　逆癌酮对 小鼠Lewis肺癌有抑癌作用,它可能是通过诱导肿瘤细胞凋亡实现的。     

局部应用内皮抑素对小鼠Lewis肺癌抑瘤作用的研究

目的我们通过局部应用内皮抑素治疗小鼠Lewis肺癌,观察内皮抑素对小鼠Lewis肺癌的抑瘤作用,及内皮抑素对VEGF表达的影响.方法用小鼠Lewis肺癌细胞种植于15只C57小鼠体内,建立小鼠Lewis肺癌移植瘤模型,当移植瘤体积生长至400～600mm3时分组并开始给药.小鼠随机分为三组,A组为空白对照组,B组每日尾静脉注射内皮抑素20μg,C组每日于肿瘤部位局部注射内皮抑素20μg,共11天.测定抑瘤率及微血管密度评价疗效,免疫组化(S-P)法测定VEGF在各组的表达水平.结果试验组较对照组抑瘤率增高(P＜0.05),微血管密度降低(P＜0.05),试验组VEGF水平较对照组低(P＜0.01).结论内皮抑素对小鼠Lewis肺癌移植瘤的生长有明显抑制作用,局部用药较全身用药的作用增强,内皮抑素可以通过降低VEGF在肿瘤组织中的表达抑制肿瘤生长.     

双氢青蒿素对Lewis肺癌小鼠的抑瘤作用

[目的]探讨双氧青蒿素对Lewis肺癌小鼠的抑瘤作用。[方法]C57BL/6J小鼠皮下接种3LL细胞（2×10^6／ml）50只．随机分为5组．分别为生理盐水组、阳性对照顺铂组、双氢青蒿素高、中、低剂量组,检测各组小鼠的体重变化和抑瘤率,瘤组织HE染色观察病理学变化,应用流式细胞术进行瘤细胞的TUNEL分析.[结果]双氢青蒿素中、高剂量组体重无生理盐水组增加明显．其抑瘤率分别为53．50％及59．24％：肿瘤组织HE染色发现：中、高剂量组瘤组织大片坏死．瘤组织内及其周围微血管少见。流式细胞术TUNEL检测结果显示,双氢青蒿素抑制肿瘤细胞增殖。[结论]口服双氢青蒿素对Lewis小鼠肺癌有明显的抑制作用;其可能是通过诱导肿瘤细胞凋亡及细胞毒作用而抑制肿瘤细胞增殖的。     

白细胞介素-18对Lewis肺癌小鼠移植瘤生长及移植瘤细胞凋亡的作用

 目的 探讨IL-18对Lewis肺癌小鼠移植瘤生长及移植瘤细胞凋亡的作用。 方法 复制Lewis肺癌小鼠模型16只,随机分为IL-18治疗组与荷瘤模型组,每组8只,分别予IL-18、生理盐水于接种第7日起每日1次腹腔注射,连续7次。观察IL-18对小鼠健康状况、移植瘤变化的影响,并用TUNEL方法检测其对移植瘤细胞凋亡的作用。 结果 IL-18对小鼠健康状况无明显影响, 对移植瘤的生长有抑制作用,肿瘤抑制率为75%,对移植瘤细胞的凋亡有促进作用。 结论 IL-18可通过促进肿瘤细胞凋亡等途径对Lewis肺癌产生抑制作用,有望成为肺癌治疗的新策略。     

20(S)-人参皂苷Rg_3对Lewis肺癌生长及转移的抑制作用

 目的观察20(S)-人参皂苷Rg3(SPG-Rg3)抗Lewis肺癌生长及转移的作用并探讨其作用机制。方法建立Lewis肺癌实体瘤模型及自发肺转移模型观察SPG-Rg3的抗肿瘤作用,并取Lewis肺癌C57BL/6荷瘤小鼠的脾脏,检测T淋巴细胞转化活性、NK细胞及IL-2活性。结果实体瘤实验中,3.0mg/kg和1.0mg/kg SPG-Rg3用药组瘤重明显低于溶剂对照组(P<0.05),0.3mg/kg组瘤重与溶剂对照组间无差异(P>0.05);SPG-Rg3用药组肺转移结节均明显少于溶剂对照组(P<0.05);SPG-Rg3用药组Lewis肺癌C57BL/6荷瘤小鼠的脾细胞对ConA和IL-2反应性以及NK活性均高于溶剂对照组(P<0.05)。结论SPG-Rg3可抑制Lewis肺癌生长及转移,可能是通过提高荷瘤小鼠的免疫功能来发挥其抑瘤作用。     

IL-12瘤苗联合放疗对小鼠Lewis肺癌作用研究

目的:建立稳定表达小鼠IL-12(Murine interleukin-12,mIL-12)的小鼠Lewis肺癌(Lewis lung carcino-ma,LLC)瘤苗LLC/mIL-12,评估瘤苗对荷鼠Lewis肺癌C57BL/6小鼠的放疗增敏作用。方法:重组质粒pcD-NA3.1( )-mIL-12用脂质体转染LLC,G418筛选后获得LLC/mIL-12瘤苗。C57BL/6小鼠皮下接种LLC细胞2×106,成瘤后将小鼠随机分成4组(n=10),第0、7、14d瘤苗组和联合组瘤内注射瘤苗,第1、3、5d放疗和联合组给予2Gy局部照射。第21d处死小鼠,观察肿瘤体积、脾细胞中CTL和NK活性以及肿瘤细胞凋亡。结果:联合组肿瘤体积显著缩小,NK和CTL活性增强,凋亡增加,与其它组相比,P<0.05;放疗组NK和CTL活性降低。结论:LLC/mIL-12瘤苗及放疗均能产生抗瘤反应,且联合组作用更强。瘤苗放疗增敏作用机制是诱导细胞凋亡,增强NK和CTL活性。     

槲皮素联合白藜芦醇对小鼠Lewis肺癌细胞生长的抑制作用

目的观察槲皮素联合白藜芦醇对小鼠Lewis肺癌细胞生长的作用及机制。方法建立C57BL/6小鼠Lewis肺癌移植瘤模型,随机分为对照组、槲皮素组、白藜芦醇组和联合用药组,每组10只,连续用药20天,于接种后第24天处死全部小鼠,比较各组肿瘤体积、肿瘤质量和抑瘤率,并用免疫组织化学、Western blot、TUNEL分别检测血管内皮生长因子 (VEGF)、基质金属蛋白酶-2(MMP-2)表达及凋亡指数(AI)。结果各用药组肿瘤的生长明显受到抑制,肿瘤体积和肿瘤质量明显低于对照组(P<0.05或0.01),联合用药组较单独用药组抑瘤作用显著增强(P<0.05)。各用药组较对照组VEGF、MMP-2表达降低(P<0.05或0.01),AI升高(P<0.05或0.01)。与单独用药组比较,联合用药组VEGF、MMP-2表达下调,AI升高,差异均有统计学意义(P<0.01)。结论槲皮素联合白藜芦醇对小鼠Lewis肺癌细胞生长具有明显的抑制作用,其机制可能与下调VEGF和MMP-2表达、促进细胞凋亡有关。     

癌得清注射液的抗癌作用

 本文用金黄色葡萄球菌(Staphylococcusaureus)、乙型溶血性链球菌(StreP－tococcushemolyticus)、卡介苗(BCG)、伤杆菌(Salmonellatyphi)等多种细菌抗原免疫SD大鼠,并从其脾脏、胸腺等淋巴组织通过透析的方法获得透析外液,再加上构才己等中药透析的小分子(MW≤10KD)物质,制备成混合制剂(癌得清)。然后以DBA/2小鼠荷L5178Y、P815肿瘤为模型,分别在肿瘤细胞接种后及接种前后腹腔注射上述制剂,观察其抑瘤作用和荷瘤小鼠脾细胞NK、LAK活性及产生IL－2活性的影响。结果表明,该制剂可明显抑制L5178Y、P815肿瘤生长并能显著增强荷瘤小鼠脾脏NK、LAK活性,增强脾细胞产生IL－2的能力。     

双氢青蒿素对Lewis肺癌小鼠的抑瘤作用及其机制探讨

目的探讨双氢青蒿素对Lewis肺癌小鼠的抑瘤效应及其作用机制。方法C57BL/6J小鼠皮下接种3LL细胞（2&#215;10^6）50只,随机分为5组,分别为生理盐水组、阳性对照顺铂组、双氢青蒿素高、中、低剂量组,检测各组小鼠的体重变化和抑瘤率,应用流式细胞术进行TUNEL分析和凋亡相关基因及其表达改变的检测。结果双氢青蒿素中、高剂量组体重较生理盐水组增加明显,其抑瘤率分别为53.5%及59.2%;流式细胞术TUNEL检测结果显示,其作用细胞凋亡和坏死同时存在。FCM法分析其对肺癌组织凋亡相关基因检测表明中、高剂量组Fas、p53等凋亡基因上调,Bcl-2抑制凋亡基因下调。结论双氢青蒿素对Lewis肺癌小鼠肺癌生长具有一定的抑制作用,能促进肿瘤细胞凋亡,其机制可能与调控细胞凋亡相关基因的表达有关。     

The mechanism of antitumor activity of total glucosides extracted from cynanchum auriculatum royle (CA)

Since the chemical structure of total glucosides from Cynanchum Auriculatum Royle (CA) is similar to that of the steroline of Marsdenia Condurago Reich, a compound which exhibits antitumor activity, research into the antitumor activity of CA was carried out. Its mechanism of action was studied in vivo with C₅₇BL/6 mice bearing Lewis lung carcinoma and in vitro, with two mouse tumor cell lines: S₁₈₀ and EAC. CA inhibited to a certain extent the growth of subcutaneously inoculated Lewis lung carcinoma and its pulmonary metastasis, and augmented the antitumor effect of cyclophosphamide. It showed a killing effect on the EAC and S₁₈₀ tumor cells of mice in vitro as well. It blocked the tumor cells of solid Lewis lung carcinoma from entering into the S stage from GI and inhibited DNA synthesis of S₁₈₀ and EAC tumor cells of mice in vitro. It also markedly increased the number of mononuclear Mφ of tumor bearing mice, stimulated the macrophagic activity of their intraperitoneal Mφ, raised the percentage of ANAE(+) lymphocytes in peripheral blood and enhanced the ABC reaction and antibody formation in tumor bearing mice.     

An oriental herbal cocktail, ka-mi-kae-kyuk-tang, exerts anti-cancer activities by targeting angiogenesis, apoptosis and metastasis

Rigorous and systematic pre-clinical studies are necessary and essential to establish the efficacy and safety of Oriental herbs and formulas in order to transform traditional herbal practices into evidence-based medicine. Here we evaluated the anti-cancer activities of the ethanol extract of Ka-mi-kae-kyuk-tang (KMKKT), a formula of ten Oriental herbs, with a battery of in vitro and in vivo mechanism-based biomarkers involving angiogenesis, apoptosis and metastasis. The results show that KMKKT suppressed the vascular endothelial responses by inhibiting basic fibroblast growth factor (bFGF)-induced ERK1/2 phosphorylation, cell migration as well as tube formation in the human umbilical vein endothelial cell model, and decreased the hypoxia-induced HIF1alpha and vascular epithelial growth factor (VEGF) expression in the mouse Lewis lung carcinoma (LLC) cells in vitro, and inhibited the bFGF-induced angiogenesis in chick chorioallantoic membrane model, and in the Matrigel plugs in mice. Intraperitoneal delivery of KMKKT potently inhibited the growth of the subcutaneously inoculated LLC cells in syngenic mice. In addition, KMKKT inhibited the invasion ability of the mouse colon 26-L5 cancer cells in vitro and decreased their formation of liver metastasis when intraportally inoculated in syngenic mice. Furthermore, KMKKT suppressed the growth of the human PC-3 prostate cancer xenografts in athymic nude mice and averted the cancer-related body weight loss. The in vivo cancer growth suppression was associated with a decreased microvessel density and VEGF abundance as well as an increased PARP cleavage and the TUNEL-positive apoptosis. Together, our data support broad-spectra in vivo anti-cancer activities of KMKKT targeting angiogenesis, apoptosis and metastasis without any adverse effect on the body weight. This formula merits serious consideration for further evaluation for the chemoprevention and treatment of cancers of multiple organ sites.     

黄芪多糖对气阴两虚Lewis肺癌荷瘤小鼠肿瘤生长、转移及细胞周期的影响

目的 观察黄芪多糖抑制气阴两虚Lewis肺癌荷瘤小鼠的生长、转移及对肺癌细胞周期的影响。方法 体外培养Lewis肺癌细胞,随机分为对照组和中药组,流式细胞术检测细胞周期;C57BL/6J小鼠90只,设空白组10只,余80只刨花烟熏并灌胃温热性的中药,移植Lewis肺癌实体肿瘤建立气阴两虚荷瘤小鼠模型并随机分为8组,比较各组小鼠抑瘤率及q值,计数外周血细胞及骨髓细胞,ELISA测定血清IL-2、IFN-γ、TNF-α、IL-10、HIF-1α、VEGF、MMP-2的含量。结果 黄芪多糖将Lewis 肺癌细胞阻滞于S期,随着药物浓度的增加,细胞凋亡逐渐增加;联合用药各组的抑瘤率高于黄芪多糖各组和顺铂组,q值均在0.85~1.15之间;与顺铂组比较,联合用药中、高剂量组小鼠外周血细胞及骨髓细胞的数量,IL-2、INF-γ、TNF-α均显著升高,IL-10、 HIF-1α、VEGF、MMP-2均显著降低（P<0.05, P<0.01）。结论 黄芪多糖在体外对Lewis肺癌细胞有抑制作用,体内与顺铂联合能抑制气阴两虚Lewis荷瘤小鼠肺癌细胞的生长、转移,提高外周血细胞和骨髓细胞的细胞数量,提高IL-2、INF-γ、TNF-α的含量,减少IL-10、HIF-1α、VEGF、MMP-2的含量。     

6-(1-Oxobutyl)-5,8-dimethoxy-1,4-naphthoquinone inhibits lewis lung cancer by antiangiogenesis and apoptosis

Lung cancer is a leading cause of cancer mortality worldwide. Novel and nontoxic agents targeting angiogenesis and tumor cell proliferation and survival are desirable for lung cancer chemoprevention and treatment. Previously we have reported that 6-(1-oxobutyl)-5,8-dimethoxy-1,4-naphthoquinone (OXO) exhibits anti-tumor activity against S-180 sarcoma in vitro and in vivo. Here we studied the anti-angiogenic and apoptogenic attributes of OXO in vitro and in vivo targeting lung cancer. In human umbilical vein endothelial cells (HUVECs), we show that OXO more potently inhibited VEGF-stimulated than basic bFGF-stimulated HUVEC proliferation and capillary differentiation. In Lewis lung carcinoma (LLC) cells, OXO not only induces S-phase arrest and mitochondria/caspase-9 pathway mediated apoptosis, but also effectively down-regulated the hypoxia-induced expression of HIF-1alpha and VEGF at mRNA and protein levels in LLC and decreased VEGF secretion into conditioned culture media. OXO significantly reduced in vivo functional angiogenesis in the mouse Matrigel plug assay. Furthermore, OXO potently inhibited the growth of LLC cells inoculated on the flank of syngenic mice at dosages that did not affect their body weight. The in vivo anti-cancer effect was associated with decreased HIF-1alpha and VEGF expression, decreased microvessel density as well as a reduction of tumor cell proliferation and increased tumor cell apoptosis. Taken together, these results demonstrate that OXO exerts anti-cancer activity through anti-angiogenesis and tumor cell cycle arrest and apoptosis. These findings warrant additional studies of OXO as a novel agent for the chemoprevention and treatment of lung cancer.     

43. Inhibition effect of Solanum nigrum. L juice on S180 ascites cancer cells

To study the action and application of Solanum nigrum.L Juice (abbreviate: S.J) on inhibiting tumors of S180 ascites cancer. Methods: Build mice tumor model through injecting S180 ascites cancer into mice's right armbet .48 male mice from KunMing of four to six weeks were randomly divided into 4 groups: Solanum Nigrum L Juice—high dosage (3mg/ml), middle dosage (1.5mg/ml), low dosage (0.75mg/ml); control group. After taking medicine for 15 days, kill the mice and measure the weight of tumor、spleen and thymus. Result: ①Tumor weights in middle and high dosage group are lighter than control group(P<0.05). ②Spleen index of test groups are different from control group(P<0.05). Conclusion: Solanum. nigrum.L Juice has inhibitory roles to S180 ascites cells.     

温阳散结汤含药血清通过NF-κB通路调控肿瘤相关巨噬细胞极化对Lewis肺癌的影响

目的:观察温阳散结汤对 Lewis 肺癌小鼠肿瘤生长及瘤组织中巨噬细胞M2型极化的影响,并探索其调控NF-κB信号通路抑制肿瘤相关巨噬细胞M2型极化,对Lewis肺癌细胞侵袭、迁移能力的影响。方法:建立C57BL/6小鼠 Lewis 肺癌移植瘤模型,温阳散结汤干预14 d 后观察小鼠瘤重和肺转移灶,计算肺转移抑瘤率,免疫组化染色检测小鼠瘤组织中巨噬细胞M2型表面标志物 CD206 的表达;温阳散结汤灌胃SD大鼠制备含药血清,采用IL-13干预RAW264.7小鼠巨噬细胞建立巨噬细胞M2型极化模型,CCK-8法检测温阳散结汤含药血清对巨噬细胞增殖的影响,流式细胞术和Western-blot检测 CD206的表达,RT-PCR检测巨噬细胞M2型标志基因的mRNA表达,ELISA法检测细胞上清中IL-1β、IL-10、TGF-β 和TNF-α 的水平变化,Western-blot检测氧化物酶体增殖物激活受体γ（PPARγ）、核因子-κB p65(NF-κB p65)和磷酸化核因子-κB p65（pNF-κB p65）的蛋白表达;收集温阳散结汤含药血清处理的M2型巨噬细胞条件培养基,将其作用于Lewis肺癌细胞,通过Transwell侵袭迁移实验检测Lewis 肺癌细胞的侵袭和运动迁移能力。结果:温阳散结汤干预后,明显减少了小鼠瘤重和肺转移灶数,并抑制了瘤组织CD206的阳性表达（P<0.05）;温阳散结汤含药血清干预后,IL-13诱导的M2型巨噬细胞中F4/80+CD206+的细胞比例和CD206蛋白表达明显减少,MRC1、Arg1、Ym1的mRNA表达水平显著降低,细胞中TNF-α、IL-1β的含量明显升高,IL-10、TGF-β的含量明显降低,同时明显下调了细胞PPARγ、NF-κB p65、pNF-κB p65的蛋白表达及pNF-κB p65/NF-κB p65比值（均P<0.05）;温阳散结汤含药血清处理的M2型巨噬细胞条件培养基干预后,Lewis肺癌细胞迁移和侵袭细胞数明显减少（P<0.05）。结论:温阳散结汤具有抑制肺癌肿瘤生长和转移的作用,其作用机制可能与调节NF-κB通路,抑制IL-13诱导的RAW264.7细胞M2型极化,从而抑制Lewis肺癌细胞侵袭和迁移能力相关。     

Gene transfer of alpha1,3-fucosyltransferase increases tumor growth of the PC-3 human prostate cancer cell line through enhanced adhesion to prostatic stromal cells

Elevated expression of sialyl Lewis X has been postulated to be a prognostic indicator of prostate cancer. However, direct evidence for the relationship between increased expression of sialyl Lewis X and malignancy of prostate cancer is still lacking. To determine whether increased levels of sialyl Lewis X leads to malignancy in prostate tumor, we transfected the human prostate cancer cell line PC-3 with alpha1,3-fucosyltransferase III (FTIII) to obtain stable transfectants, PC-3-FTIII lines, that highly express sialyl Lewis X. When inoculated in the prostate of nude mice, PC-3-FTIII cells produced large prostate tumors, while mock-transfected PC-3 cells, which are negative for sialyl Lewis X antigen, produced small prostate tumors. The aggressive tumor formation by PC-3-FTIII cells was inhibited by preincubation of the tumor cells with anti-sialyl Lewis X antibody, by the presence of sialyl Lewis X oligosaccharide or by selectin ligand mimic peptide but not by control peptide. PC-3-FTIII cells and mock-transfected PC-3 cells exhibited no significant difference in cell numbers when cultured in vitro. Remarkably, PC-3-FTIII adhered to prostatic stromal cells in vitro with higher affinity than mock-transfected PC-3. Such adhesion was inhibited by preincubation of PC-3-FTIII cells with antisialyl Lewis X antibody, by the addition of sialyl Lewis X oligosaccharide or by selectin ligand mimic peptide. However, anti-E-selectin, anti-P-selectin or anti-L-selectin antibodies did not inhibit the adhesion of PC-3-FTIII cells to the stromal cells. These results suggest that prostate cancer cells gain aggressiveness through adhesive interaction with prostatic stromal cells by a novel mechanism involving sialyl Lewis X.     

地塞米松对小鼠H_(22)肿瘤生长及血管内皮生长因子表达的影响

 目的　探讨地塞米松对小鼠H2 2 肿瘤生长及血管内皮生长因子 (VEGF)表达的影响。方法　BALB c小鼠皮下接种H2 2 肿瘤细胞 ,腹腔注射地塞米松 ,每天测肿瘤直径。用抗CD31单抗对肿瘤组织做免疫组化微血管计数。噻唑蓝法检测地塞米松对体外培养的H2 2 细胞和人脐静脉内皮细胞ECV 30 4增殖的影响。RT PCR法测定肿瘤组织和体外培养细胞的VEGFmRNA表达。结果　地塞米松可以显著抑制体内H2 2 肿瘤的生长 ,给药组肿瘤体积与对照组比较P <0 .0 5。给药组微血管计数和VEGFmRNA表达比对照组显著减少。体外实验中 ,随着地塞米松浓度的增加 ,ECV 30 4细胞的增殖率降低 ;H2 2 细胞中VEGFmR NA表达下降。结论　地塞米松可以显著抑制小鼠H2 2 肿瘤的生长。地塞米松对体内H2 2 肿瘤组织和体外H2 2 细胞的VEGFmRNA表达均有显著的抑制作用。地塞米松对肿瘤细胞VEGF表达的抑制作用可能是其抗H2 2 肿瘤及抗血管生成的一个重要原因。     

依维莫司负调控mTOR信号通路对膀胱癌BTT739细胞增殖和迁移的抑制作用及机制研究

目的:探索mTOR抑制剂依维莫司(RAD001)对膀胱癌BTT739细胞在体外和小鼠体内增殖和迁移的抑制作用及其作用机制,为依维莫司在临床上治疗膀胱癌提供理论依据。方法:体外培养小鼠膀胱癌BTT739细胞,待其生长至对数期后用梯度浓度的RAD001（0 nmol/L、5 nmol/L、10 nmol/L和20 nmol/L）干预24 h,用MTT法检测细胞增殖的变化;Tranwell实验检测RAD001对BTT739细胞迁移的影响;ELISA试剂盒检测细胞分泌蛋白E-钙黏蛋白表达的变化。Western blot检测Akt、mTOR、4EBP和PTEN蛋白的表达,RT-PCR检测mTOR mRNA的表达。将BTT739细胞接种于昆明鼠皮下,待成瘤后,用依维莫司干预14天,取小鼠肿瘤组织检测E-钙黏蛋白及AKT、mTOR、4EBP和PTEN蛋白表达的情况。结果:体外实验显示RAD001可以抑制BTT739细胞的增殖和迁移并与剂量有关（P＜0.05）。还可以增加E-钙黏蛋白的含量,抑制癌细胞的转移。实验证明RAD001在体外抑制Akt、mTOR、4EBP的表达,促进肿瘤抑制因子PTEN的表达（P＜0.05）。将BTT739细胞接种于小鼠皮下后发现,RAD001在小鼠体内亦可以抑制病变情况,促进PTEN和E-钙黏蛋白的表达,抑制Akt、mTOR和4EBP的表达。结论:mTOR抑制剂依维莫司通过抑制mTOR信号通路抑制膀胱癌细胞BTT739的增殖、迁移和侵袭。     

中肺合剂对S180肉瘤、Lewis肺癌荷瘤小鼠抑瘤作用实验研究

[目的]探讨中肺合剂对S180肉瘤、Lewis肺癌荷瘤小鼠的抑瘤作用,并初步探索其机制.[方法]采用S180肉瘤、Lewis肺癌移植瘤小鼠模型,分荷瘤阴性对照组、低、中、高剂量中肺合剂组和CTX阳性对照组,观察中肺合剂对两种荷瘤小鼠肿瘤生长的抑制作用;运用EnVision免疫组化染色法观察中肺合剂对Lewis肺癌荷瘤小鼠肿瘤组织cvclinD1周期蛋白表达的影响。[结果]中肺合剂低、中、高剂量组和CTX组对移植性荷S180小鼠的抑瘤率分别为16．48％、29．73％、23．51％、41．87％;与阴性对照组相比,中肺合剂低、中、高剂量组和CTX组瘤重均有显著性差异（P〈0．05）。中肺合剂低、中、高剂量组和CTX组对移植性荷Lewis肺癌小鼠的抑制率分别为22．33％、39．02％、25．48％、64．16％;与阴性对照组相比,低剂量组和高剂量组瘤重均有显著性差异（P〈0．05）,中剂量组和CTX化疗组瘤重均有非常显著性差异（P〈0．01）。中肺合剂能下调Lewis肺癌小鼠肿瘤组织cyclinD1蛋白的表达水平,中、高剂量组阳性表达率与阴性对照组相比,有显著性差异（P〈0．051：[结论]中肺合剂对S180和Lewis荷瘤小鼠有明显的抑瘤作用,量-效关系呈抛物线型．其抑瘤作用可能与下调肿瘤组织cyclinD1蛋白表达水平有关。