Metastatic human hepatocellular carcinoma models in nude mice and cell line with metastatic potential

Metastatic human HCC model is needed for the studies on mechanism and intervention of metastatic recurrence. By using orthotopic implantation of histologically intact tissues of 30 surgical specimens, a patient-like metastatic model of human HCC in nude mice (LCI-D20) and a low metastatic model of human HCC in nude mice (LCI-D35) have been established. All mice with transplanted LCI-D20 tumors exhibited extremely high metastatic ability including spontaneous metastasis to liver, lungs, lymph nodes and peritoneal seeding. Remarkable difference was also found in expression of some of the invasiveness related genes and growth factors between the LCI-D20 and LCI-D35 tumors. PAI-1 increased gradually following tumor progression in LCI-D20 model, and correlated with tumor size and AFP level. Phasic expression of tissue intercellular adhesion molecule-1 in this model was also observed. Using corneal micropocket model, it was demonstrated that the vascular response induced by LCI-D20 tumor was stronger than that induced by LCI-D35 tumor. Similar report on metastatic human HCC model in nude mice and human HCC cell line with metastatic potential was rarely found in the literature. This LCI-D20 model has been widely used for the studies on intervention of metastasis, including anti-angiogenesis,antisense approach, metalloproteinase inhibitor, differentiation inducer, etc. It is concluded that the establishment of metastatic human HCC model in nude mice and human HCC cell line with metastatic potential will provide important models for the in vitro and in vitro study of HCC invasiveness, angiogenesis as well as intervention of HCC recurrence.

Effects of epidermal growth factor on the growth of human gastric cancer cell and the implanted tumor of nude mice

AIM: Epidermal growth factor (EGF) plays an important role in the regulation of gastrointestinal tissue growth and development, and it can stimulate epithelial proliferation, cell differentiation and growth. It has been established that the EGF can promote gastric cytoprotection and ulcer healing. But the potential ability of EGF to regulate the gastric cancer growth is unknown. This study is to investigate the influence of EGF on human gastric cancer cell and the implanted tumor growth of nude mice. METHODS: The cell growth rates of human gastric adenocarcinoma cell lines MKN-28, MKN-45, SGC-7901 and normal human gastric epithelial cells 3T3 were assessed when incubated with recombinant human EGF (rhEGF, 0.05, 0.1, 0.5, 1.0, 10, 50, 100 mg.L(-1)) using MTT method. The cells of MKN-28, MKN-45, SGC-7901 (gastric cancer tissue 1.5mm(3)) were implanted in the BALB/cA nude mice for 10 days.The EGF was given intraperitoneally (15, 30, 60 microg.kg(-1)) for 3 weeks. The body weights of the tumor-bearing animals and their tumor mass were measured afterwards to assess the mitogenic effect of rhEGF in the nude mice. RESULTS: Within the concentration range of 0.05-100mg.L(-1), rhEGF could increase the cell growth of normal 3T3 cells (cell growth rate 100% vs 102.8%, P【0.05), but partially restrain the gastric cancer cell growth. The latter effect was related to cell differentiation. In 15-60 microg/kg rhEGF groups, the mean implanted tumor mass of MKN-28 cell were 1.75 g, 1.91 g, 2.08 g/NS group 1.97 g (P】0.05), the mean tumor mass of SGC-7901 cell were 1.53 g, 1.07 g, 1.20 g/NS group 1.07 g (P】0.05), and for MKN-45 cell, the tumor mass were respectively 1.92 g, 1.29 g, 1.77 /NS group 1.82 g (P】0.05). So rhEGF had no obvious effect on implanted MKN-28, SGC-7901 and MKN-45 tumor growth. CONCLUSION: EGF has no stimulating effect on the human gastric cancer cell growth neither in vitro nor in vivo.

RNA interference of pax2 inhibits growth of transplanted human endometrial cancer cells in nude mice

The development of human endometrial carcinoma(HEC) is a complex pathologic process involves several oncogenes and tumor suppressor genes.The full-length paired-box gene 2(pax2),a recently discovered oncogene,promotes cell proliferation and growth and inhibits apoptosis of HEC cells.Here,we examined the effect of pax2 small interfering RNA(siRNA) on the growth of transplanted HEC cells in nude mice.The expression of Pax2 in 21 cases of normal endometrium and 38 cases of HEC was examined by immohistochemistry(IHC).HEC models were developed by subcutaneously transferring HEC cells into nude mice,followed by treatment with empty lentivirus vector,lentivirus vector-based pax2 siRNA,and phosphate buffered saline,respectively.Four weeks later,tumor size was measured,tumor inhibition rate was calculated,and histological analyses were conducted after staining with hematoxylin and eosin.The expression of Pax2 and Bcl-2 was detected by Western blot;proliferating cell nuclear antigen(PCNA) was detected by IHC.Significant differences were observed in the positive rate of Pax2 between normal endometrium and HEC(14.2% vs.60.5%,P<0.01).The expression index of Pax2 in well differentiated tumors was 1.88±1.68,much lower than that in tumors of moderate(3.07±1.96,P<0.05) or poor differentiation(5.45±2.76,P<0.01).Tumor necrosis increased,nuclear basophilia stain decreased,tumor growth was inhibited,and PCNA,Pax2,and Bcl-2 expression was reduced in HEC models treated with pax2 siRNA.These results indicate that Pax2 expression is related to HEC tumor biology with the increased expression of Pax2 correlated to malignancy.pax2 siRNA down-regulates Pax2 expression and inhibits tumorigenesis of HEC in nude mice,possibly due to cell apoptosis and the inhibition of tumor proliferation induced by down-regulation of Bcl-2.

Orthotopical transplantation of human renal carcinoma tissue into nude mice and the establishment of a high metastatic cell line MRCC

Objective To establish a SOI model of human renal carcinoma and a high metastatic cell subline. Methods A human renal cell line RCC-9863 has been established by inoculating a human renal tumor tissue into nude mice s. c.. When RCC-9863 passaged for 20 times, the tissue from the same xemotransplant tumor were used to construct SOI model. Cultured the metastatic tissue in vitro, the tumor cell suspension was then injected orthotopically, The metastatic tissue obtained underwent the same procedure again. At last, the metastatic tumor was cultured in vitro and cloned. Results 15 days later, a tumor mass sized 1. 7 cm × 0. 6 cm in the nude mouse’s renal parenchyma was grown which lobulated, rude, and with multiply blood vessels and 55 days later later the mouse became moribund and metastases in the lungs were formed. The transplanted renal tumor in the SOI model grew fast and invasively and metastasized to lungs, lymphatic node and liver. A subline, MRCC, with metastatic ability to the lung was selected.

Antitumor activities of human autologous cytokineinduced killer(CIK)cells against hepatocellular carcinoma cells in vitro and in vivo

AIM: To characterize the anticancer function of cytokine-induced killer cells (CIK) and develop an adoptive immunotherapy for the patients with primary hepatocellular carcinoma (HCC), we evaluated the proliferation rate, phenotype and the antitumor activity of human CIK cells from healthy donors and HCC patients in vitro and in vivo. METHODS: Peripheral blood mononuclear cells (PBMC) from healthy donors and patients with primary HCC were incubated in vitro and induced into CIK cells in the presence of various cytokines such as interferon-gamma (IFN-gamma), interleukin-1 (IL-1), IL-2 and monoclonal antibody (mAb) against CD3. The phenotype and characterization of CIK cells were identified by flow cytometric analysis. The cytotoxicity of CIK cells was determined by (51)Cr release assay. RESULTS: The CIK cells were shown to be a heterogeneous population with different cellular phenotypes. The percentage of CD3+/CD56+ positive cells, the dominant effector cells, in total CIK cells from healthy donors and HCC patients, significantly increased from 0.1-0.13% at day 0 to 19.0-20.5% at day 21 incubation, which suggested that the CD3+ CD56+ positive cells proliferated faster than other cell populations of CIK cells in the protocol used in this study. After 28 day in vitro incubation, the CIK cells from patients with HCC and healthy donors increased by more than 300-fold and 500-fold in proliferation cell number, respectively. CIK cells originated from HCC patients possessed a higher in vitro antitumor cytotoxic activity on autologous HCC cells than the autologous lymphokine-activated killer (LAK) cells and PBMC cells. In in vivo animal experiment, CIK cells had stronger effects on the inhibition of tumor growth in Balb/c nude mice bearing BEL-7402-producing tumor than LAK cells (mean inhibitory rate, 84.7% vs 52.8%, P【0.05) or PBMC (mean inhibitory rate, 84.7% vs 37.1%, P【0.01). CONCLUSION: Autologous CIK cells are of highly efficient cytotoxic effector cells against primary hepatocellular carcinoma cells and might serve as an alternative adoptive therapeutic strategy for HCC patients.

Establishment of an orthotopic transplantation tumor model of hepatocellular carcinoma in mice

AIM:To improve the outcome of orthotopic transplantation in a mouse model,we used an absorbable gelatin sponge(AGS) in nude mice to establish an orthotopic implantation tumor model.METHODS:MHCC-97L hepatocellular carcinoma(HCC)cells stably expressing the luciferase gene were injected into the subcutaneous region of nude mice.One week later,the ectopic tumors were harvested and transplanted into the left liver lobe of nude mice.The AGS was used to establish the nude mouse orthotopic implantation tumor model.The tumor suppressor gene,paired box gene 5(PAX5),which is a tumor suppressor in HCC,was transfected into HCC cells to validate the model.Tumor growth was measured by bioluminescence imaging technology.Semi-quantitative reverse transcription polymerase chain reaction(RT-PCR) and histopathology were used to confirm the tumorigenicity of the implanted tumor from the MHCC-97L cell line.RESULTS:We successfully developed an orthotopic transplantation tumor model in nude mice with the use of an AGS.The success rate of tumor transplantation was improved from 60% in the control group to 100% in the experimental group using AGS.The detection of fluorescent signals showed that tumors grew in all live nude mice.The mice were divided into 3 groups:AGS-,AGS+/PAX5-and AGS+/PAX5 +.Tumor size was significantly smaller in PAX5 transfected nude mice compared to control mice(P < 0.0001).These fluorescent signal results were consistent with observations made during surgery.Pathologic examination further confirmed that the tissues from the ectopic tumor were HCC.Results from RT-PCR proved that the HCC originated from MHCC-97L cells.CONCLUSION:Using an AGS is a convenient and efficient way of establishing an indirect orthotopic liver transplantation tumor model with a high success rate.

Experimental study on antitumor effect of arsenic trioxide in combination with cisplatin or doxorubicin on hepatocellular carcinoma

INTRODUCTIONThe main component of a traditional Chinese drug 'Pishuang'. arsenic trioxide (As2O3), has obviously selective anti-tumor effect on human hepatocellular carcinoma (HCC)in both in vitro and in vivo studies[1-5]. Due to limited effectiveness when any anti-carcinogen is used alone and obviously increased toxicity when the dose is raised, there is no exception for As2O3. Furthermore, combined chemotherapy contributes to improve therapeutic effectiveness, disperse toxicity and surmount drug-resistance,in which the combination of traditional Chinese and modern medicine has more advantages and characteristics. As a result,we made an experimental study on anti-tumor effect of As2O3in combination with cisplantin (PDD) or doxorubicin (ADM)on HCC. to investigate the possibility of AS2O3 in combination with PDD or ADM and nature of interaction between them,and to provide experimental basis for clinical application.

Human breast carcinoma xenografts in nude mice

OBJECTIVE: To investigate spontaneous metastasis, micrometastasis and genetic stability in human breast carcinoma xenografts in nude mice. METHODS: Intact tissue from surgical specimens from breast carcinoma patients was xenografted into nude mice and transplanted from generation to generation. Cells from the xenografts were cultured in vitro and retransplanted into nude mice. Microsatellite DNA in the genome of human breast carcinomas, xenotransplanted tumors and metastases in nude mice were analyzed at three microsatellite loci. RESULTS: The tumorigenicity of orthotopic xenotransplantation was 88.6% (31/35), with a metastatic rate of 41.9% (13/31). Cells from xenotransplants were successfully cultured in vitro. The taking rate of retransplantation into nude mice and the spontaneous lung metastasis rate were both 100% (10/10). Microsatellite DNA sequences in the genome of xenotransplanted tumors and metastases in nude mice were identical with that of the original human breast carcinoma at three microsatellite loci. CONCLUSIONS: Tumorigenicity and metastatic potential can be improved in human breast carcinoma xenografts using intact fresh tumor tissue and orthotopic grafts. Xenotransplanted tumors and tumors after serial passage maintained the genetic stability. The detection of microsatellite DNA may identify micrometastases in a nude mouse model.

Antihepatoma effect of alpha-fetoprotein antisense phosphorothioate oligodeoxyribonucleotides in vitro and in mice

AIM: To evaluate antihepatoma effect of antisense phosphorothioate oligodeoxyribonucleotides (S-ODNs) targeted to alpha-fetoprotein (AFP) genes in vitro and in nude mice. METHODS: AFP gene expression was examined by immunocytochemical method or enzyme-linked immunosorbent assay. Effect of S-ODNs on SMMC-7721 human hepatoma cell growth in vitro was determined using microculture tetrazolium assay. In vitro antitumor activities of S-ODNs were monitored by measuring tumor weight differences in treated and control mice bearing SMMC-7721 xenografts. Induction of cell apoptosis was evaluated by fluorescence-activated cell sorter (FACS) analysis. RESULTS: Antisense S-ODN treatment led to reduced AFP gene expression. Specific antisense S-ODNs, but not control S-ODNs, inhibited the growth of hepatoma cells in vitro. In vitro, only antisense S-ODNs exhibited obvious antitumor activities. FACS analysis revealed that the growth inhibition by antisense S-ODNs was associated with their cell apoptosis induction. CONCLUSION: Antisense S-ODNs targeted to AFP genes inhibit the growth of human hepatoma cells and solid hepatoma, which is related to their cell apoptosis induction.

Transcription factor EGR-1 inhibits growth of hepatocellular carcinoma and esophageal carcinoma cell lines

AIM: The transcription factor EGR-1 (early growth response gene-1) plays an important role in cell growth, differentiation and development. It has identified that EGR-1 has significant transformation suppression activity in some neoplasms, such as fibrosarcoma, breast carcinoma. This experiment was designed to investigate the role of egr-1 in the cancerous process of hepatocellular carcinoma (HCC) and esophageal carcinoma (EC), and then to appraise the effects of EGR-1 on the growth of these tumor cells. METHODS: Firstly, the transcription and expression of egr-1 in HCC and EC, paracancerous tissues and their normal counterpart parts were detected by in situ hybridization and immunohistochemistry, with normal human breast and mouse brain tissues as positive controls. Egr-1 gene was then transfected into HCC (HHCC, SMMC7721) and EC (ECa109) cell lines in which no egr-1 transcription and expression were present. The cell growth speed, FCM cell cycle, plate clone formation and tumorigenicity in nude mice were observed and the controls were the cell lines transfected with vector only. RESULTS: Little or no egr-1 transcription and expression were detected in HCC, EC and normal liver tissues. The expression of egr-1 were found higher in hepatocellular paracancerous tissue (transcription level P=0.000; expression level P=0.143, probably because fewer in number of cases) and dysplastic tissue of esophageal cancer (transcription level P=0.000; expression level P=0.001). The growth rate of egr-1-transfected HHCC (HCC cell line) cells and ECa109 (EC cell line) cells was much slower than that of the controls. The proportion of S phase cell, clone formation and tumorigenicity were significantly lower than these of the controls' (decreased 45.5% in HHCC cells and 34.1% in ECa109 cells; 46.6% and 41.8%; 80.4% and 72.6% respectively). There were no obvious differences between SMMC7721 (HCC) egr-1-transfected cells and the controls with regard to the above items. CONCLUSION: The decreased expression of egr-1 might play a role in the dysregulation of normal growth in the cancerous process of HCC and EC. Egr-1 gene of transfected HHCC and ECa109 cells showed obvious suppression of the cell growth and malignant phenotypes, but no suppression in SMMC7721 (HCC cell line) cells.

Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97

AIM: To establish clone cells with different metastatic potential for the study of metastasis-related mechanisms. METHODS: Cloning procedure was performed on parental hepatocellular carcinoma (HCC) cell line MHCC97, and biological characteristics of the target clones selected by in vivo screening were studied. RESULTS: Two clones with high (MHCC97-H) and low (MHCC97-L) metastatic potential were isolated from the parent cell line. Compared with MHCC97-L, MHCC97-H had smaller cell size (average cell diameter 43 microm vs 50 microm) and faster in vitro and in vivo growth rate (tumor cell doubling time was 34.2h vs 60.0h). The main ranges of chromosomes were 55-58 in MHCC97-H and 57-62 in MHCC97-L. Boyden chamber in vitro invasion assay demonstrated that the number of penetrating cells through the artificial basement membrane was (37.5 +/- 11.0) cells/field for MHCC97-H vs (17.7 +/- 6.3)/field for MHCC97-L. The proportions of cells in G0-G1 phase, S phase, and G2-M phase for MHCC97-H/MHCC97-L were 0.56/0.65, 0.28/0.25 and 0.16/0.10, respectively, as measured by flow cytometry. The serum AFP levels in nude mice 5wk after orthotopic implantation of tumor tissue were (246 +/- 66) microg.L(-1) for MHCC97-H and (91 +/- 66) microg.L(-1) for MHCC97-L. The pulmonary metastatic rate was 100% (10/10) vs 40% (4/10). CONCLUSION: Two clones of the same genetic background but with different biological behaviors were established, which could be valuable models for investigation on HCC metastasis.

Reduction of tumorigenicity of SMMC-7721 hepatoma cells by vascular endothelial growth factor antisense gene therapy

AIM: To test the hypothesis to block VEGF expression of SMMC-7721 hepatoma cells may inhibit tumor growth using the rat hepatoma model. METHODS: Amplify the 200 VEGF cDNA fragment and insert it into human U6 gene cassette in the reverse orientation transcribing small antisense RNA which could specifically interact with VEGF165, and VEGF121 mRNA. Construct the retroviral vector containing this antisense VEGF U6 cassette and package the replication-deficient recombinant retrovirus. SMMC-7721 cells were transduced with these virus and positive clones were selected with G418. PCR and Southern blot analysis were performed to determine if U6 cassette integrated into the genomic DNA of positive clone. Transfected tumor cells were evaluated for RNA expression by ribonuclease protection assays. The VEGF protein in the supernatant of parental tumor cells and genetically modified tumor cells was determined with ELISA. In vitro and in vivo growth properties of antisense VEGF cell clone in nude mice were analyzed. RESULTS: Restriction enzyme digestion and PCR sequencing verified that the antisense VEGF RNA retroviral vector was successfully constructed.After G418 selection, resistant SMMC-7721 cell clone was picked up. PCR and Southern blot analysis suggested that U6 cassette was integrated into the cell genomic DNA. Stable SMMC-7721 cell clone transduced with U6 antisense RNA cassette could express 200 bp small antisense VEGF RNA and secrete reduced levels of VEGF in culture condition. Production of VEGF by antisense transgene-expressing cells was 65+/-10 ng/L per 10(6) cells, 42045 ng/L per 10(6) cells in sense group and 485+/-30 ng/L per 10(6) cells in the negative control group, (P【 0.05). The antisense-VEGF cell clone appeared phenotypically indistinguishable from SMMC-7721 cells and SMMC-7721 cells transfected sense VEGF. The growth rate of the antisense-VEGF cell clone was the same as the control cells. When S.C. was implanted into nude mice, growth of antisense-VEGF cell lines was greatly inhibited compared with control cells. CONCLUSION: Expression of antisense VEGF RNA in SMMC-7721 cells could decrease the tumorigenicity, and antisense-VEGF gene therapy may be an adjuvant treatment for hepatoma.

Retrovirus-mediated herpes simplex virus thymidine kinase gene therapy approach for hepatocellular carcinoma

The therapeutic effect of herpes simplex virus thymidine kinase/ganciclovir (HSV-tk/GCV) system on hepatocellular carcinoma was studied in this experiment. The tk-containing retroviral recombinants were used to infect hepatoma cells (BEL-7402) and the cells were treated with ganciclovir (0-1000 microg/ml). The results showed that HSV-tk gene could be efficiently transferred in vitro into hepatoma cells and stably expressed. The growth potential of the tk-containing cells was significantly inhibited by GCV (P

蛇葡萄素对人肺癌GLC-82裸鼠移植瘤的抑制作用

目的 :为了进一步研究蛇葡萄素体内抗肿瘤作用 ,我们进行了人肺癌GLC 82裸小鼠移植瘤的抑瘤实验研究。方法 :人肺癌GLC 82细胞接种于裸鼠腋窝建立移植模型。荷瘤BALB/C裸鼠随机分成 6组 ,蛇葡萄素以 3个剂量腹腔给药。观察肿瘤体积、相对肿瘤体积、瘤重、相对肿瘤增殖率以及肿瘤生长曲线 ,以此评价蛇葡萄素的抗瘤作用。结果 :2 5 0mg/kg蛇葡萄素对裸鼠移植瘤的抑制率 ,二次实验结果分别为 35 5 % (P <0 0 1)和 37 1% (P<0 0 1) ,相对肿瘤增殖率则分别为 5 5 2 4 % (P <0 0 1)和 5 7 71% (P <0 0 5 )。结论 :蛇葡萄素对人肺癌GLC 82裸鼠移植瘤具有显著的抑制作用。

益气化瘀解毒方对人肝癌裸鼠HepG2移植瘤MVD、HIF1a、VEGF/KDR表达的影响

目的:观察益气化瘀解毒方对人肝癌裸鼠HepG2移植瘤MVD、HIF1a、VEGF/KDR表达的影响。方法:建立人肝癌移植瘤模型,分为7组,分别以益气化瘀解毒方、黄芪、莪术、重楼、壁虎、顺铂药物干预21 d后,取瘤组织行免疫组化检测。结果:①对MVD的影响:全方组与除顺铂外的其它组比较有统计学意义(P<0.05);除黄芪、莪术外的其它组与空白组比较有统计学意义(P<0.05)。②对HIF1a的影响:全方组与除黄芪外的其它组比较有统计学意义(P<0.05);各组与空白组比较有统计学意义(P<0.05)。③对VEGF表达的影响:全方组与除顺铂外的其它组比较均有统计学意义(P<0.05);莪术组与空白组比较无统计学意义(P>0.05)。④对KDR表达的影响:全方组与其它各组比较均有统计学意义(P<0.05);黄芪组与空白组比较无统计学意义(P>0.05)。结论:益气化瘀解毒方能明显抑制人肝癌裸鼠移植瘤MVD、HIF1a、VEGF和KDR的表达而发挥抗血管生成作用;各单味药对其作用各有侧重,但作为整方的有机组成从不同环节发挥了协同作用。

三氧化二砷抗裸鼠人结肠癌移植瘤作用及其机制的研究

目的:探讨三氧化二砷(As2O3)抗人结肠腺癌裸鼠移植瘤作用、作用机制以及药物毒性。方法:建立人结肠癌裸鼠移植瘤模型,随机分为4组,即生理盐水组、5-氟脲嘧啶(5-FU)组、低剂量As2O3组和高剂量As2O3组,比较各组的抑瘤作用和裸鼠一般状态的变化,并对标本分别行光镜和电镜观察、原位末端标记(TUNEL)、免疫组化和血常规检测。结果:As2O3能够明显抑制结肠癌裸鼠移植瘤的增长,并能诱导癌细胞凋亡,调控相关基因表达;未见As2O3引起明显肝、肾和造血系统损害。结论:As2O3对人结肠腺癌细胞裸鼠移植瘤具有显著的抗癌作用,此作用可能与诱导癌细胞凋亡密切相关,且受到多种有关基因的调控;但是对裸鼠的肝、肾和造血系统无明显的毒性。

蛇葡萄素的抗肿瘤作用研究

目的:研究蛇葡萄素在体内外对肿瘤生长的抑制作用。方法:用MTT法测定蛇葡萄素对人鼻咽癌HK-1细胞和人乳腺癌MCF-7细胞的体外细胞毒实验;观察蛇葡萄素对C57BL/6小鼠移植性B16黑色素瘤的体内抑瘤作用;用流式细胞仪测定含药小鼠血清对B16细胞周期及细胞增殖的影响。结果:蛇葡萄素对人鼻咽癌HK-1细胞及人乳腺癌MCF-7细胞的IC50分别为50.0μg/ml及79.4μg/ml。在100mg/kg、150mg/kg及200mg/kg的剂量下,与生理盐水对照组比较,蛇葡萄素对C57BL/6小鼠移植性B16黑色素瘤的抑瘤率分别为25.54%(P<0.05)、32.03%(P<0.01)及54.55%(P<0.001);60mg/kg氮烯咪胺(阳性对照组)的抑瘤率为59.31%,与200mg/kg的药物组比较无显著性差异(P>0.05);在150mg/kg及200mg/kg蛇葡萄素腹腔给药后10min的小鼠含药血清的作用下,B16细胞的G1期和G2/M期细胞数增高,S期细胞数减少,分裂增殖指数分别降低19.1%及21.7%。结论:蛇葡萄素对体外肿瘤细胞HK-1和MCF-7的增殖,及体内小鼠移植性B16黑色素瘤的生长均具有显著的抑制作用。

兴安升麻总苷抗肿瘤药效研究

目的观察升麻总苷对小鼠移植性肿瘤和人肿瘤细胞裸鼠移植瘤的体内抗肿瘤作用,以及体外对人肿瘤细胞株的抑瘤活性,并对其抑瘤机制进行初步探讨。方法MTT法检测体外升麻总苷对人肿瘤细胞的增殖抑制作用;体内实验观察其对小鼠S180和裸鼠体内移植人肺腺癌A549的抑制作用;同时采用流式细胞仪和肿瘤病理切片对接种的A549肿瘤细胞凋亡进行观察。结果升麻总苷对A549、HepG2、HL60、Eca-109、MDA-MB231肿瘤细胞的半数抑制浓度(IC50)分别为20.3、27.1、21.2、23.4和32.7μg/mL。小鼠口服升麻总苷100mg/kg和200mg/kg可明显抑制S180移植瘤(抑瘤率分别为42.8%和54.6%)和裸鼠移植人肺腺癌A549的生长(T/C值分别为58.1%和52.2%),肿瘤组织病理切片和流式细胞仪对A549肿瘤细胞凋亡的检测显示,升麻提取物可诱导体内肿瘤细胞凋亡。结论升麻体内体外均具有抗肿瘤活性,其体内抑制肿瘤生长可能与诱导细胞凋亡相关。

双氢青蒿素对人结直肠癌裸鼠移植瘤的抑制作用及机制研究

目的研究双氢青蒿素(Dihydroartemisinine,DHA)对人结直肠癌LoVo细胞裸鼠移植瘤的抑制作用及可能机制。方法将人结直肠癌LoVo细胞接种于Balb/c裸鼠皮下,建立结直肠癌模型。将造模成功裸鼠随机分为5组,即模型对照组,阳性对照组(5-Fu 20 mg/kg),DHA高、中、低剂量组(120,60,30 mg/kg)。观察各组动物肿瘤的重量及抑瘤率;采用免疫组化SABC法测定各组裸鼠肿瘤组织B-cell lymphoma-leukemia-2 gene(Bcl-2)和血管内皮生长因子(VEGF)的表达。结果 5-Fu组及DHA高、中剂量组的抑瘤率显著高于模型组(P<0.01或P<0.05);5-Fu组和DHA高剂量组Bcl-2的阳性表达率显著低于模型对照组(P<0.01或P<0.05);5-Fu组VEGF的阳性表达率显著低于模型对照组(P<0.05)。结论双氢青蒿素具有较强的抗结直肠癌活性,其作用机制可能是抑制抗凋亡基因Bcl-2的表达,从而刺激结直肠癌细胞凋亡。

E1A基因对裸鼠移植瘤生长抑制及其初步作用机制的实验研究

目的:探讨E1A基因对人宫颈癌细胞裸鼠移植瘤生长的抑制作用及其相关作用机制。方法:通过裸鼠移植瘤实验,观察E1A基因对人宫颈癌细胞裸鼠移植瘤生长的的抑制作用。采用免疫组织化学染色检测E1A基因对肿瘤细胞凋亡及Survivin基因和Caspase-3基因表达的影响。结果:裸鼠移植瘤实验结果显示:Hela-E1A细胞形成的肿瘤较Hela和Hela-vect的出瘤时间晚,生长慢,瘤重小,抑瘤率分别为83.42%和84.74%。免疫组织化学染色显示:E1A基因组细胞凋亡数量较Hela和Hela-vect组明显增多,Caspase-3基因(凋亡促进因子)呈高表达,Survivin基因(调亡抑制因子)的表达明显降低。结论:E1A基因能够明显抑制人宫颈癌细胞裸鼠移植瘤的生长,该作用可能与E1A基因激活Caspase-3基因和降低Survivin基因的表达有关。