人肾透明细胞癌细胞系RCC-9863的建立及其生物学特性

目的　建立一株新的人肾癌细胞系 ,为肾癌的基础研究提供实验模型。　方法　由肾癌手术标本中切取 1.0cm× 1.0cm组织块 ,分别包埋于 3只裸鼠右后肢皮下 ,连续传代 3次 ,取移植瘤体外培养。　结果　 2只裸鼠皮下移植瘤生长 ,继续传代 ,肿瘤生长速度明显加快。取移植瘤标本体外培养得到肾癌细胞系RCC 986 3(简称R)。形态结构 ,分化程度与原发瘤一致 ,染色体形态仍为人类核型 ,众数为 6 2～ 6 8,占 75 %。细胞周期分析G1期 6 3.8% ,G2期 11.4 % ,S1期 2 4 .8% ,细胞群体倍增时间为 37.7h。　结论　通过裸鼠皮下移植瘤建立的肾癌细胞系RCC 986 3与原发癌保持相同的生物学特性 ,体外连续传代 1年以上 ,传代 110代。细胞形态不变 ,生长周期恒定 ,已成为一个稳定的细胞系     

利用裸鼠建立人前列腺肿瘤原位移植模型

目的：应用前列腺梭形细胞瘤细胞系PC－98106建立人前列腺肿瘤原位移植模型，观察肿瘤局部浸润和远处转移情况。方法：8只裸鼠取下腹部切口，显露膀胱和前列腺，将PC－98106细胞悬液0．2－0．4ml注射于前列腺后叶包膜下。观察裸鼠生命体征的变化，取原位肿瘤、膀胱、盆腔淋巴结、肺脏、肝脏、脾脏标本福马林液固定48h，苏木素－伊红染色后显微镜下观察。原位移植瘤电镜观察，X线照射显示骨骼转移情况。结果：5只模型建立成功，10d左右，可触及前列腺部位有小结节。60d左右，裸鼠出现明显恶液质。解剖后发现盆腔淋巴结肿大，显微镜观察膀胱有浸泣，肝脏有微转移灶。X线未见骨骼转移。结论：前列腺肿瘤原位移植模型的转归接近于临床，是肿瘤转移研究的较好模型。     

ACHN人肾癌裸鼠移植瘤模型的生物学特性及放射治疗观察

目的观察ACHN人肾癌裸鼠皮下移植瘤模型的生物学特性及其对放射线的敏感性。方法分别利用瘤细胞悬液接种法、瘤块包埋法获得ACHN人肾癌裸鼠移植瘤模型，观察移植瘤的生长情况、成瘤率、肝肺转移率及病理形态。ACHN皮下荷瘤鼠14只，随机分为空白对照组和放疗组，6MV X直线加速器对放疗组移植瘤局部单次照射12Gy，观察裸鼠的耐受性及肿瘤生长抑制情况；透射电镜观察瘤细胞超微结构变化。结果瘤块包埋法成瘤稳定，成瘤率92．5%，肝转移率43％，未见肺转移。裸鼠对12Gy射线局部照射耐受良好，放疗组移植瘤与空白对照组比较生长明显受抑制（P〈0．01）；放疗组瘤细胞凋亡现象明显。结论ACHN人肾癌裸鼠移植瘤模型可作为肾癌放疗实验研究的有效工具。     

p53引起的细胞凋亡和恶性肿瘤的基因治疗

肿瘤抑制基因p53的基因治疗可抑制大多数p53突变的恶性肿瘤细胞的体外增殖和在裸鼠上的成瘤能力，对于原位和皮下的肿瘤移植模型，WT-p53基因治疗可使肿瘤缩小。这些变化与调节细胞周期和肿瘤细胞凋亡有关。     

bFGF及其抗体对移植前列腺癌生长影响的研究

目的探讨碱性成纤维细胞生长因子（bFGF）及其抗体对裸鼠人前列腺癌移植瘤生长的影响和作用机制。方法建立人前列腺癌细胞系（DU-145）细胞裸鼠皮下移植瘤动物模型,观察移植肿瘤生长及荷瘤裸鼠状况,观察bFGF与bFGF抗体对裸鼠皮下瘤生长状况的影响;应用免疫组织化学方法检测肿瘤组织中增殖细胞核抗原（PCNA）和CD34的表达。结果 1bFGF治疗组移植瘤重量和体积分别比对照组显著增多,移植瘤组织中PCNA指数和微血管密度（MVD）也显著提高;2bFGF抗体治疗组移植瘤体积重量和体积比对照组均显著减少,移植瘤组织中PCNA指数和MVD也显著降低。结论 bFGF能促进前列腺癌细胞的增殖生长,bFGF抗体则能抑制肿瘤的细胞生长,其作用机制与调控肿瘤细胞的增殖和血管形成密切相关。     

p53引起的细胞凋亡和恶性肿瘤的基因治疗：文献综述

肿瘤抑制基因Ｐ５３的基因治疗可抑制大多数Ｐ５３突变的恶性肿瘤的体外增殖和在裸鼠上的成瘤能力，对于原位和皮下的肿瘤移植模型下ＷＴ－Ｐ５３基因治疗可使肿瘤缩小。     

重组人生长激素对裸鼠人胃癌移植瘤细胞周期的影响

目的：研究重组人生长激素（recombinant human growth hormone．rhGH）对裸鼠人胃癌移植瘤细胞周期的影响。方法：将24只实验裸鼠随机分为对照组、顺铂（DDP）组、rhGH组和DDP＋rhGH组，每组6只，将体外培养的人胃癌细胞MKN45接种于裸鼠右臀部皮下，裸鼠胃癌移植瘤模型成功建立后开始连续给药6d，给药结束后处死裸鼠。取出移植瘤，用流式细胞仪检测细胞周期，并分析细胞周期、细胞增殖指数（PI）及DNA抑制率。结果：给药结束后DDP＋rhGH组和DDP组S期细胞明显减少，而rhGH组和对照组比较增多（P〈0．05）；G0-G1期、G2-M期的细胞增殖指数（PI）和DNA抑制率各组间比较差异均无统计学意义（P〉0．05）。rhGH组细胞凋亡率增加而对照组、DDP组和DDP＋rhGH组减少（P〈0．05）。结论：未发现rhGH对裸鼠人胃癌移植瘤细胞有明显促进分裂增殖作用。     

人胆管癌裸鼠移植瘤模型的建立

目的：建立人胆管癌裸鼠移植瘤1号和2号模型。方法：将人胆管癌组织接种于裸鼠皮下和肝脏，逐代观察移植瘤的生长情况，绘制其生长曲线，进行形态学和生物学特性鉴定。结果：建立了人胆管高分化粘液腺癌裸鼠移植瘤1号和中分化乳头状腺癌裸鼠移植瘤2号模型。皮下移植瘤生长率为40％；1、2号模型移植成功率分别为97．7％和100％，潜伏期分别为26d和217d。移植瘤在形态和生物学上仍保持人胆管癌的特点。结论：裸鼠移植瘤1、2号模型是一种接近人体的胆管癌模型，可为胆管癌研究提供实验平台。     

人胃癌细胞原位移植裸鼠原发灶与肝转移灶体外化疗药敏性差异

目的：比较人胃癌原位移植裸鼠肝转移模型原发灶、肝转移灶肿瘤细胞对化疗药物敏感性。 方法：将裸鼠皮下传代的SGC-7901细胞株实体瘤组织块移植于裸鼠胃壁,建立人胃癌裸鼠原位移植模型,待其发生肝转移后取胃原发灶、肝转移灶肿瘤细胞,SRB法检测肿瘤细胞对氟尿嘧啶（5-FU）,顺铂（CDDP）,奥沙利铂（L-OHP）,表阿霉素（eADM）,丝裂霉素（MMC）,长春新碱（VCR）,氨甲喋呤（MTX）7种化疗药物的体外敏感性。 结果：成功建立裸鼠原位移植胃癌转移模型,肿瘤原位移植成瘤率100%,肝转移率75%;7种药物中L-OHP,VCR对原发灶肿瘤细胞的抑制率高于肝转移灶,而eADM,MMC对肝转移灶肿瘤细胞的抑制率高于原发灶（均P<0.05）;5-FU,L-OHP,MTX对原发灶与肝转移灶的抑制率具正相关性（r=0.5203;0.4424;0.3851,均P<0.05）。 结论：胃癌原位移植动物的原发灶和肝转移灶细胞的对化疗药物药敏性存在差异,以原发灶药敏检测结果指导针对肝转移灶的治疗可能是不准确的。     

骨肉瘤高转移细胞SOSP—M1生物学稳定性及组织学起源的鉴定

目的 检测人骨肉瘤细胞高转移亚系SOSP-M1在裸小鼠体内传代过程中的生物学稳定性并鉴定其组织学特性。方法 利用原位移植将细胞系在33只裸鼠体内连续传代,组织块培养收集各代肺转移灶的肿瘤细胞。观察各代肿瘤细胞致瘤率、转移率、形态结构、骨形成蛋白、波形蛋白,肌动蛋白、神经元特异性烯醇化酶的表达情况以及遗传学方面的变化。结果 各代肿瘤细胞致瘤率均为100%,体内增殖稳定,肺转移率在80%以上,其显微及超微结构形态,抗原表达、染色体数目及结构变化均符合人骨肉瘤的特征。结论 该细胞亚系在裸鼠体内传代生物学特性相对稳定,是人骨肉瘤实验研究的良好模型。     

Herbal therapy PC-SPES: in vitro effects and evaluation of its efficacy in 69 patients with prostate cancer

PURPOSE: We investigate the potential use of the phytotherapeutic PC-SPES to treat human prostate cancer, and evaluate its in vivo and in vitro activity, and clinical efficacy. MATERIALS AND METHODS: PC-SPES was evaluated for its ability to induce apoptosis on prostate cancer cell lines LNCaP, PC3 and DU145. The effect of oral PC-SPES on growth of PC3 tumors present in male immunodeficient mice was studied. A total of 30 male nude mice were divided in 5 groups. In groups 1 control and 2 full dose therapy was started the same day of the tumor injection. In groups 3 control, 4 half dose and 5 full dose PC-SPES therapy was initiated 1 week after tumor injection. A total of 69 patients with prostate cancer were treated with 3 capsules of 320 mg. PC-SPES daily. Serum prostate specific antigen (PSA) responses and side effects were evaluated. RESULTS: All of the cultured prostate cancer cell lines had a significant dose dependent induction of apoptosis following exposure to an alcoholic PC-SPES extract. Immunodeficient mice xenografted with the PC3 cell line had reduced tumor volume compared with sham treated controls when they were treated with a PC-SPES extract from the time of tumor cell implantation (931 +/- 89 versus 1,424 +/- 685 mm.3, p not significant) but not when the treatment was begun 1 week after tumor cell implantation. The testis, prostate, bladder and seminal vesicles of the treated mice were significantly reduced in weight compared with the sham treated animals. Of the patients with prostate cancer 82% had decreased serum PSA 2 months, 78% 6 months and 88% 12 months after treatment with PC-SPES. Side effects in the treated patient population included nipple tenderness in 42% and phlebitis requiring heparinization in 2%. CONCLUSIONS: An extract of the phytotherapeutic agent PC-SPES proved to be active in inducing apoptosis of hormone sensitive and insensitive prostate cancer cells in vitro, and in suppressing the growth rate of a hormone insensitive prostate cancer cell line in vivo. The overwhelming majority of patients with prostate cancer treated with the agent experienced a decrease in serum PSA but also demonstrated a side effect profile comparable to estrogen treatment.     

Chemotherapy of carcinoma of the prostate and testis: experimental study in vivo and in vitro

Prostate cancer: Considering the stagnation in chemotherapy of prostate cancer in recent years, the following experiments were carried out to determine their clinical value. Surgical specimens from 6 patients, 2 permanent cell lines (EB 33 and PC 93) originated from human prostate cancer and a tumor line serially transplanted in nude mice (PC-NCC) were subjected to chemosensitivity tests such as human tumor cloning assay (HTCA) and/or in vivo tumor growth curve experiments using nude mice. The possible chemosensitive drugs screened by using surgical specimens and PC-NCC tumor were cisplatinum (CDDP), bleomycin (BLM), 5-FU, vincristine (VCR), adriamycin (ADM) and methotrexate (MTX). Most of these drugs were also judged as "effective" by HTCA using a permanent cell line. The minimal discrepancy among them may lead to the conclusion that an in vitro assay using a cell line can substitute for the assay using surgical specimens which can not be obtained frequently. Partly based on the data obtained a chemotherapy regimen, VPM-CisCF, consisting of VCR, peplomycin, MTX, CDDP, cytosine arabinoside (Ara-C) and 5-FU, was designed. The effectiveness of this regimen was demonstrated experimentally. Testis cancer: Two different lines of experiments were performed. A human testicular cancer serially transplanted in nude mice was repeatedly exposed to CDDP in vivo to obtain hyposensitivity to this drug. The synergistic effect of CDDP and VP-16 was demonstrated in the tumor thus obtained. One of its mechanisms has been suggested by partial accumulation of cancer cells in the G1-S and G2-M phase in which CDDP exerts its potential effect.(ABSTRACT TRUNCATED AT 250 WORDS)     

人食管鳞癌细胞系RJEC-2的建立及其生物学特性分析

目的:建立新的人食管鳞状细胞癌细胞系并分析其生物学特性,为食管癌分子机制和治疗干预的研究提供新的实验模型。方法:采用组织块培养法,从病人食管癌组织中分离纯化鳞状上皮癌细胞并建立细胞系。对细胞系的形态特点、细胞角蛋白表达、生长特征、细胞周期分布、细胞遗传特征和致瘤能力进行了研究分析。结果:建立了食管鳞状细胞癌细胞系RJEC-2,已在体外持续传代4个多月,传至46代,生长稳定;该细胞系具有鳞状上皮细胞的形态和特点:单层贴壁生长,免疫组化显示细胞角蛋白表达阳性,电镜下可见明显的胞质内张力纤维束和细胞间桥粒;群体倍增时间为46.5 h,细胞培养至90%汇合时,细胞周期分析显示G0/G1期平均占56.72%,S期平均占33.96%,G2/M期平均占9.32%;细胞染色体结构和数量异常,呈现肿瘤细胞特性;细胞呈克隆性生长,平板克隆平均形成率为13.93%,裸鼠移植瘤实验表明细胞具有致瘤能力,病理分析显示移植瘤与病人肿瘤病理形态特征相似,均为中、高分化鳞状细胞癌。结论:成功建立的人食管鳞癌细胞系RJEC-2,为食管癌发病机制和治疗方案的研究提供了新的研究实验模型。     

来源于转移灶的人肝细胞癌细胞系HN-HC1的建立和特性

目的建立新的转移性人肝细胞癌细胞系并研究其生物学特性。方法从人肝细胞癌的腹腔转移灶取材,将标本分离成单细胞悬液,使用培养液进行原代和传代培养,取名为HN-HC1细胞,观察细胞的形态,绘制细胞生长曲线。于裸小鼠腹腔接种第18代HN-HC1细胞2×106个,观察癌细胞的生物学特性,检测AFP的表达。结果 HN-HC1细胞体外连续传代至第18代,形态上具有典型的恶性上皮细胞的特征,裸小鼠腹腔HN-HC1细胞成瘤率100%,移植瘤细胞中AFP呈强阳性表达。结论 HN-HC1细胞可能成为较稳定的来源于转移灶的人肝细胞癌细胞系,HN-HC1裸小鼠移植瘤是一种较理想的肝细胞癌动物模型,为肝细胞癌的研究提供了良好的动物实验平台。     

ErbB-4 may control behavior of prostate cancer cells and serve as a target for molecular therapy

PURPOSE: To assess ErbB-4 expression in advanced human prostate cancer (PC) cell lines, the role of ErbB-4 in motility, migration, and proliferative/tumorigenic potential of PC cells, and efficacy of anti-ErbB-4 monoclonal antibody (Mab) treatment on PC cells in vitro and tumor growth in vivo. MATERIALS AND METHODS: Established advanced human PC cell lines (PC-3, Cl-1, and Du-145) were evaluated for ErbB-4 expression. Several Cl-1 cell line clones expressing various levels of ErbB-4 were isolated, their motility, migration capacity, and in vitro proliferation as well as survival following Mab treatment were evaluated. Tumorigenicity and proliferation capacity of these clones in vivo and efficacy of Mab treatment on tumor growth were estimated by measurements of subcutaneous tumors developed in nude mice. RESULTS: PC cell lines studied express ErbB-4. Both PC-3 and Du-145 cell lines express high ErbB-4 levels; only 50% of Cl-1 cells express ErbB-4 with large heterogeneity. Cl-1 sub-clones highly expressing ErbB-4 showed increased cell motility, migration, and proliferation rate in vitro and enhanced growth in vivo, compared to clones with low ErbB-4 expression. Mab treatment inhibited the growth of cells expressing high but not low ErbB-4 levels in vitro and decreased the growth of subcutaneous tumors in nude mice generated by ErbB-4 highly expressing cells. CONCLUSIONS: High expression of ErbB-4 in prostate cancer Cl-1 cell clones correlated with high proliferative and migration capacity and high tumorigenic potential. The inhibitory effect of Mab on cell proliferation and on subcutaneous tumor growth suggests ErbB-4's potential as a target for molecular anticancer therapy.     

Influence of pertussis toxine on local progression and metastasis after orthotopic implantation of the human prostate cancer cell line PC3 in nude mice

Tumor cell migration is a fundamental process of metastasis. Pertussis toxine inhibits lysophosphatidic acid related cell migration by ADP-ribosylation of G proteins. We examined the influence of pertussis toxine (PTX) on progression and metastasis of the human hormone-insensitive prostate cancer cell line PC-3 after orthotopic implantation in nude mice. In 30 athymic male nude mice (NMRI) 5x10(5) PC-3 cells were injected into the dorsal prostate. After 7 d 15 mice received a total of six intraperitoneal injections of 5 micro g PTX/100 g body weight at an interval of 4 d. The other 15 mice received phosphate buffered saline and served as control. All mice were killed at 37 d followed by macroscopical and histological evaluation of local tumor growth and metastasis. In the control group tumorigenicity was 100% (15 out of 15). Mean weight of the tumor bearing unit of prostate and seminal vesicles was 541 mg (243-763 mg). The rate of positive lymphnodes was 100% with a mean transversal diameter of 3.9 mm (1.2-5.4 mm). In the PTX group local take rate was 100% with a mean weight of 251 mg (88-478 mg) (P two sided <0.0001). The rate of positive lymphnodes was 60% (9 out of 15) (P=0.017) with a mean transversal diameter of 2.3 mm (1.0-4.5 mm). PTX following orthotopic implantation of the human hormone-insensitive PC-3 cell line significantly reduces local tumor growth as well as metastasis to locoregional lymphnodes.     

Contragestazol （DL111-IT） inhibits proliferation of human androgen-independent prostate cancer cell line PC3 in vitro and in vivo

Aim： To evaluate the antiproliferative activity of contragestazol （DL111-IT） on the human prostate cancer cell line PC3 in vitro and in vivo and to elucidate its potential molecular mechanisms. Methods： The cell killing ability of DL111-IT was measured by the 3-（4,5-dimethylthia-zol,2-yl）-2,5-diphenyltetrazolium bromide （MTT） reagent assay method and the tumor xenograft model. The cell cycle was analyzed by flow cytometry and protein expression, including retinoblastoma （pRb）, cyclin-dependent kinase 4 （CDK4） and cyclin D 1, was detected by Western blotting. Results： DL111-IT exhibited high efficiency on cell growth inhibition of the human androgen-independent prostate cancer cell line PC3. The drug concentration that yielded 50 % cell inhibition （IC50 value） was 9.9 mg/mL. In the PC3 tumor xenograft study, DL111-IT （1.25 mg/kg-20.0 mg/kg） given once a day for 10 days significantly inhibited tumor growth, with the inhibition rate ranging from 21% to 50 %. Flow cytometric analysis indicated that DL111-IT could cause GI arrest in the PC3 cell line, but not apoptosis. DL111-IT enhanced pRb expression and down-regulated CDK4 and cyclin D 1 expression, suggesting that cell cycle regulation might contribute to the anticancer property of DL 111- IT. Conclusion： DL111-1T inhibits the proliferation of human androgen-independent prostate cancer cell line PC3 in vitro and in vivo by a cell cycle regulation pathway.     

沉默PAK6基因可抑制前列腺癌生长并增强多烯紫杉醇的作用

目的 观察沉默p21活化激酶6(PAK6)基因对前列腺癌生长及联用多烯紫杉醇的作用.方法 转染siRNA(30 nmoL/L)前列腺癌细胞PC-3和DU145中,检测细胞增殖、集落形成能力、侵袭性能力、细胞周期及凋亡.以3 x 106个PC-3细胞(100μl)注射裸鼠皮下成瘤,观察siRNA及联用多烯紫杉醇对肿瘤生长效应.结果 PAK6-siRNA能有效沉默PAK6并抑制前列腺癌细胞的生长、集落形成能力和侵袭力、使细胞周期停滞在G2/M期.还可抑制裸鼠体内移植瘤生长.PAK6-siRNA治疗组瘤体重为(146.0±8.5)mg,低于对照组(451.0±22.3)mg(P<0.01).PAK6-siRNA在体内外均可增强多烯紫杉醇作用.结论 沉默PAK6可抑制前列腺癌生长并增强多烯紫杉醇的化疗作用.