The recruitment of exogenous endothelial progenitor cells in lung tumor model of nude mice

Background and Objective: Endothelial progenitor cells (EPCs) play an important role in hypoxia-triggered tumor vasculogenesis. However, the homing of exogenous EPCs in tumors is still unclear. In this study, we investigated the recruitment of exogenous EPCs in human lung adenocarcinoma model of nude mice. Methods: EPCs labeled with green fluorescence protein (GFP) were transplanted into nude mice bearing human lung adenocarcinoma. The growth of tumor was observed. After the mice were killed, GFP-EPCs in different tissues were examined by fluorescence. The tumor tissues were stained for CD133, hypoxia-inducible factor-1alpha (HIF-1α), stromal cell-derived factor-1α (SDF-1α), and vascular endothelial growth factor receptor (KDR). Real-time polymerase chain reaction of CD133, HIF-1α, SDF-1α, and VEGF-1 were also performed. Results: The growth of tumor in EPC group was significantly faster than that in saline solution group (P < 0.05). Under fluorescence microscope, GFP-EPCs were strongly expressed in both tumor and bone marrow. EPCs were recruited to the tumor periphery to participate in tumor vasculogenesis. The expression of CD133, HIF-1α, and SDF-1 mRNA in tumor and bone marrow were significantly higher than that in the liver, spleen, and skin (P < 0.05). Conclusions: Exogenous EPCs can be recruited to tumor and accelerate tumor growth. Except tumor, bone marrow can also recruit EPCs.

Immuno-protective effect of tumor cell vaccine on Kunming mice bearing Ehrlich ascites tumor

AIM To evaluate the immunity of chemically modified tumor cell vaccine.METHODS Tumor cell vaccines (TCV) were prepared by incubating the live Ehrlich ascites tumor cells with concanavalin A-mitomycin C (ConA-MMC), mitomycin C (MMC), concanavalin A-glutaraldehyde (ConA-Glu), glutaraldehyde (Glu), or paraformaldehyde (Para), respectively. The whole cell or soluble forms of the vaccines were administered intraperitoneally into Kunming mice once a week for three times prior to the intraperitoneal inoculation of a lethal dose of live tumor cells. A second challenge with live tumor cells was given four weeks later. Survival and antibody production of the mice were analyzed.RESULTS After the first challenge, the mice, received whole TCV of ConA-MMC, MMC (P＜0.01) and Glu (P＜0.05) promoted survival incidence than the controls. All the treated mice had the survival time prolonged. ConA-MMC vaccine treated mice had longer survival days than that of ConA-Glu ones (P＜0.05). For the soluble TCV immunized mice, those treated with vaccines of Para (P＜0.01), ConA-Para and ConA-Glu (P＜0.05) had longer survival periods compared with that of the controls. Following the second challenge, survival incidence of the mice received vaccines of ConA-MMC, MMC, ConA-Glu or Glu was significantly increased (P＜0.01). Moreover, all the treated mice had the survival time prolonged, and ConA-MMC vaccine treated mice had longer survival days than that of Para treated ones (P＜0.05). Antibodies against Ehrlich ascites tumor cells were found to be positive in sera of the mice treated with whole TCV of ConA-MMC.CONCLUSION Ehrlich ascites tumor cells are immunogenic when treated with ConA-MMC, MMC, ConA-Glu, Glu or Para, which might act as safe and effective tumor vaccines with safety and effectiveness.

Functional Mechanism of Resveratrol in Inhabiting Growth of Cells ls174t and Its Mechanism in Subcutaneously Transplanted Tumor of Nude Mice

To explore the functional mechanism of Resveratrol against colon cancer cells ls174t and the growth of colon cancer tissue of tumor-bearing mice, MTT method was used to observe the functions of resveratrol for inhibition against cells ls174t in vitro. Transmission electron microscope was used to observe the cell apoptosis. FCM assay was performed to measure the change of the cell apoptosis rate and of cell cycle. RT-PCR method was used to detect the expressions of bcl-2 and bax mRNA. Western blot method was used to detect the expressions of bcl-2 and bax protein. Ceils ls174t were transplanted subcutaneously to nude mice to observe the effect of resveratrol on the growth of subcutaneously transplanted tumor, RT-PCR method was used to detect the expressions of bcl-2 and bax mRNA in the tumor tissue. Western blot method was used to detect the expressions of bcl-2 and bax protein in the tumor tissue. Resveratrol has an effect of inhibiting proliferation of cells ls174t in vitro（P〈0.01）. It is able to induce the apoptosis of cells ls174t, causing the decrease in the expression of bcl-2 and the increase in the expression of bax. Resveratrol could inhibit the growth of subcutaneously transplanted tumor of nude mice（P〈0.05）, causing the decrease in the expression of bcl-2 and the increase in the expression of bax. Resveratrol can inhibit the growth of cells 174t and the growth of subcutaneously transplanted tumor. The mechanism is possibly related to the induction of the cell apoptosis and the regulation of bcl-2/bax expression.

Effect of Recql5 deficiency on the intestinal tumor susceptibility of Apc^(min) mice

AIM:To investigate whether Recql5,a DNA helicase that plays an important role in the maintenance of genome integrity,is a tumor suppressor in the gastrointestinal tract in mice.METHODS:We generated cohorts of both Recql5-proficient and Recql5-deficient Apcmin/+mice and compared the tumor susceptibility in their gastrointestinal tracts.RESULTS:Recql5 deficiency in Apcmin/+mice resulted in a significant increase in the tumor incidence in both the colon(P=0.0162)and the small intestine(P<0.01).These findings have provided the first genetic evidence for a tumor suppression role of Recql5 in the gastrointestinal tract of mice.Importantly,since mouse Recql5 and human RECQL5 are highly conserved,these findings also suggest that RECQL5 may be a tu-mor suppressor for human colon cancer.CONCLUSION:Recql5 has a tumor suppression role in the mouse gastrointestinal tract.

Anti-tumor Effect and Mechanism of Pratia Extract on H22 Tumor-bearing Mice

[Objectives]This study was conducted to investigate the inhibitory effect of pratia extract on H22 tumor-bearing mice and the effects on immune organs.[Methods]With the application of H22 liver tumor-bearing mice as an animal model,the animals were divided into such three Pratia extract groups as the high,medium and low dose groups(400,200 and 100 mg/kg)and cyclophosphamide CTX group(20 mg/kg).15 d after the administration,the animals were killed by cervical dislocation,and the tumors,thymuses and spleens were taken and weighed,followed by the calculation of the tumor inhibitory rate and the thymus and spleen index,and the serum tumor necrosis factor-α(TNF-α)and interleukin-2(IL-2)levels were determined by ELISA assay.[Results]The inhibitory rates were 54.1,32.6 and 8.2%,respectively,and there were significant differences from the model group(P<0.05);and the spleen index of the tumor-bearing mice was reduced,while the thymus index was improved.The serological results showed that the drug-administrated groups significantly improved the IL-2 levels in the tumor-bearing mice,but had no effects on TNF-α.[Conclusions]Pratia extract has an antitumor effect on H22 tumor-bearing mice,and show certain dose-effect relationship,and its mechanism may be related to enhancing the immune function in tumor-bearing mice by regulating IL-2.

Evaluation of Tumor Formation of Three Bladder Cancer Cell Lines in Nude Mice

This study examined the differences in tumor formation of three bladder tumor cell lines （BIU-87, T24 and EJ） after subcutaneously transplanted into nude mice, in order to find the best technique for establishing in vivo bladder tumor model. BIU-87, T24 and EJ cells at logarithmic phase were re-suspended in serum-free medium. The cells suspensions of the identical concentration were subcutaneously transplanted into nude mice and then the success rate and tumor growth were compared among the three cell groups. The results of tumor formation were pathologically evaluated. Lung, liver and kidney tissues were also pathologically examined for distant metastasis. The proliferation of the three cells were determined by immunohistochemically detecting the PCNA expression in the tumors. The results showed that the success rates of EJ and T24 cells were significantly higher than that of BIU-87 cells and no distant metastasis was noted among the three groups. The proliferation levels of EJ and T24 cells was significantly higher than that of BIU-87. But at the later stage of tumor formation, as compared with T24 cells, EJ grew more vigorously, soon resulting in the central necrosis of tumor, which affected the measurement of the actual size of the tumors. Moreover, PCNA staining exhibited that the proliferation of EJ and T24 was significantly higher than that of BIU-87 cells. It is concluded that as compared with BIU-87 cells, EJ and T24 cells had higher success rates, with not significant differences in death rate and distant metastasis found among them. There existed no significant difference in tumor formation between EJ and T24 cells and T24 cells do not rupture easily, which makes it a better cell line for the establishment of in vivo bladder tumor model.

Anti-tumor effect of LTA combined with 5-FU on H22 tumor bearing mice

Objective: To study the effect of lipoteichoic acid(LTA) and 5-FU on the expression of caspase-3, EGFR, TGF-α proteins of tumor tissue of H22 cancer bearing mice and its antitumor mechanism. Methods: A total of 40 SPF grade Kunming mice were selected to establish H22 liver cancer model, and then the mice were divided into 4 groups at random with ten mice in each group. Group A was given saline lavage treatment, Group B was treated with 5-FU by intraperitomeal injection, Group C was treated with LTA by lump body injection; Group D was treated with LTA by lump body injection and 5-FU by intraperitomeal injection. Two weeks after the treatment, the mice in each group were executed and the tumor tissue was stripping and weighted, and the tumor growth inhibition ratio was calculated. Then the tumor tissue was processed for conventional embedding, sectioned to observe the expression of caspase-3, EGFR, TGF-α by immunohistochemical staining method. Results: The tumor inhibitory rate o f Group D was significantly higher than Groups B and C(P<0.05); B, the tumor inhibitory rate o f Group B had no statistical difference compared with Group C(P>0.05). The IDO values of TGF-α, EGFR proteins in Groups B, C, D mice tumor tissue were significantly lower than that in group A(P<0.05); while IDO value of caspase-3 in Groups B, C, D group mice tumor tissue was significantly higher than that in Group A(P<0.05). The IDO value of TGF-α, EGFR in Group D mice tumor tissue were significantly lower than that in Groups B and C; While IDO value of aspase-3 in Group D was significantly higher than that in Groups B and C(P<0.05). Conclusions: LTA combined with 5-FU can effectively inhibit the tumorigenesis of H22 tumor bearing mice, increase the caspase-3 protein expression, inhibit TGF –α and EGFR protein expression, further promote tumor cell apoptosis and play a synergistic antitumor effect.

Immunotherapeutic effects of dendiritic cells vaccine pulsed with tumor cell lysate in mice with pancreatic carcinoma

Objective To observe the immunotherapeutic effects of dendritic cells vaccine pulsed with tumor cell lystate on mice with pancreatic carcinoma. Methods Dendritic cells (MTSC4) were pulsed with tumor cells lysate. The immune preventative and immnotherapeutic effects of DC vaccines on mice with pancreatic carcinoma were assessed. Results After vaccination of the DC vaccines,mice remained tumor-free for at least 25 days in DCs vaccines group,but in other groups the subcutaneous implantation tumorigenesis were found beginning 3 to 9 days. CTL stimulated by DC vaccines effected cytolytic activity against pancreatic carcinoma cells. The survival period was obviously prolonged in DCs vaccines group (56 ±9)d than in other groups P【0.01) and tumors (1.4 ±0.8)g in DCs vaccines group were significantly smaller than that in other groups (P 【 0. 05). Conclusion Tumor cell lysate-pulsed dendrtic cells vaccines can induce a specific and effective immune response against pancreatic carcinoma cell implanted in mice.

Effect of wheat maxibastion of Guanyuan point on immune function and antitumor effect in mice with tumor

Objective To investigate wheat maxibastion of Guanyuan point on immune function and antitumor effect in mice with tumor.Method Group experiment and statistical analysis were conducted.Results were analyzed comparatively.Result Wheat maxibastion of Guanyuan point could reduce ATEP(associated tumor erythrocyte rate) and direct tumor erythrocyte rate(DTER) and raise excitotumorous erythrocyte rate(ETER)(P< 0.05,P< 0.01) and stimulate the reduced activity of NK cells and IL 2 significantly(P< 0.01) in mice with tumor.Conclusion Wheat maxibastion of Guanyuan point can increase the adhesion ability to tumor cells of RBC,and increase the activity of NK cell and IL 2.

Gene expression and cellular localizations of tumor necrosis factor-α at the site of implanted bovine cancellous bone in mice

The objective of this study was to determine if mRNA encoding for tumor necrosls factor-α(TNFα) was present at the site of implanted bovine cancellous bone and to observe the cellular localizations. The particles of bovine cancellous bone treated by special chemical reagents were implanted in the mouse’s muscle pouch. removed 5.10 and 20 days after implantation, and the specimens were processed for determining the expression and cellular localizations of TNFα mRNA, which was performed by a nonradioactive in situ hybridization technique. The results showed that (1) 5, 10 and 20 days after transplantation, the TNFα mRNA expressions were positive, andthe positive rate of expression was the highest by 10 days (P<0. 05 ). (2)There was strong hybridization signal localization to the nuclei of morphologically ldentifiable monocytes and multinucleated giant cells. (3)Similar activity was detected in the cytoplasm and (or) nuclei of partial adjacent mesenchymal cells, fibroblasts as well as striated muscle fibers. This finding tended to indicate that mRNA encoding for TNFα was intensely expressed in several kinds of cells and that TNFα seemed to be of importance for the modulation of local cellular immunity in the region of implanted xenogeneic bone.

Anti-breast cancer properties and toxicity of Dillenia suffruticosa root aqueous extract in BALB/c mice

Objective:To determine the anti-breast cancer activities and the safety oral consumption of Dillenia suffruticosa root aqueous extract(DRAE)in BALB/c mice.Methods:In the anti-breast cancer study,female BALB/c mice were divided into five groups(n=12),which were(1)positive control(with breast cancer,untreated),(2)negative control(without breast cancer,untreated)and other three groups of mice with breast cancer treated with 1 000,500 and 250 mg/kg of DRAE,respectively,by oral gavage for 28 days.All mice except from the negative control group were injected into the mammary fat pad with 4T1 cells(1×1054T1 cells/0.1 m L of phosphate buffer solution).DRAE was administered orally on Day 11 after the tumor has developed.Results:The tumor volume of the 1 000 mg/kg of DRAE group reduced significantly compared to the positive control while treatment with 500 mg/kg of DRAE had significantly inhibited metastasis to the heart.In the acute toxicity study,treatment with up to5 000 mg/kg of DRAE was not toxic to the animals,indicating its safety when a large amount of this plant extract was ingested.Based on the sub-acute toxicity study,treatment of the highest dose of DRAE(1 000 mg/kg)had mild liver toxicity indicated by mild focal hemorrhage.Conclusions:DRAE possesses anti-breast cancer properties but at the same time it shows mild toxicity to the liver.The non observable adverse effect dose for DRAE is500 mg/kg.

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.

Inhibition of KIT RNAi mediated with adenovirus in gastrointestinal stromal tumor xenograft

AIM: To investigate a therapeutic method for gastrointestinal stromal tumor (GIST) based on KIT RNA interference (RNAi) with AdMax adenovirus. METHODS: KIT short hairpin RNA (shRNA), whose lateral sides were decorated with restriction endonuclease sequences, was designed. T 4 DNA ligase catalyzed the joint of the KIT shRNA and the green fluorescent protein-containing PDC316-EGFP-U6 to form PDC316EGFP-U6-KIT. Homologous recombination of AdEGFPU6-KIT was performed with the AdMax system. Heterotopically transplanted GISTs were established in nude mice. AdEGFP-U6-KIT was intratumorally injected. The volume, inhibition ratio of tumor and CD117 expression of GIST graft tumor in nude mice were compared between test and control groups. RESULTS: The length of KIT shRNA was determined to be about 50bp by agarose electrophoresis. Gene se-quencing detected the designed KIT RNAi sequence in PDC316-EGFP-U6-KIT. After transfection with AdEGFPU6-KIT, 293 cells displayed green fluorescence. The physical and infective titers of AdEGFP-U6-KIT were 5 × 10 11 viral particles/mL and 5.67 × 10 7 plaque forming units/mL, respectively. The mean volume of the grafted tumor was significantly smaller in test mice than in control mice (75.3 ± 22.9 mm 3 vs 988.6 ± 30.5 mm 3 , t = -18.132, P < 0.05). The inhibition ratio of the tumors was 59.6% in the test group. CD117 positive expression was evident in two cases (20%) in the test group and 10 cases (100%) in the control group (χ 2 = 10.2083, P < 0.005). CONCLUSION: AdEGFP-U6-KIT is successfully constructed, and KIT RNAi mediated with Admax vector system can effectively inhibit the expression of the KIT gene and the growth of GIST in nude mice.

A Totally Absorbable Multilayer PLGA Implant Device Containing Doxorubicin Inhibited Tumor Growth and Metastasis without Systemic Toxicity in Murine Breast Cancer and an Ideal Pharmacological Paradigm for Regional Chemotherapy

We hypothesize that a cylinder implant made of multilayer Poly-lactic-co-glycolic-acid (PLGA) membrane can be a method for controlled and extended drug release. We fashioned a multilayer cylindrical implant termed STID100 that released doxorubicin for 3 weeks in an orthotopic 4T1 breast cancer model in Balb/C mice. This implant starts as a thin doxorubicin-embedded PLGA membrane, and is then rolled into a cylinder containing an air gap between the membrane layers. Its controlled sustained release delivered 2× the amount of the intravenous (IV) equivalent of doxorubicin, inhibited the primary tumor, and prevented lung metastasis. Importantly it did not cause weight loss, splenomegaly, or cardiac toxicity vs systemically administrated doxorubicin. This favorable safety profile is further substantiated by the finding of no detectable plasma doxorubicin in multiple time points during the 3-week period, and low tumor doxorubicin concentration. The implant system delivered to the specification of an ideal pharmacological paradigm might offer a better coverage of the local tumor, significantly preventing metastatic spread with less drug toxicity to many vital organs, compared to the traditional pharmacology of IV route. The profile of STID made it an attractive therapeutic alternative in metastatic tumor prevention, pain management and many other diverse clinical scenarios.

基于TGF-β1/CD147信号探讨慢性束缚应激促小鼠乳腺癌进展及逍遥散调节机制研究

目的基于TGF-β1/CD147信号探讨慢性束缚应激促小鼠乳腺癌进展及逍遥散调节机制。方法40只BABL/c小鼠随机分为移植瘤组(Tumor)、模型组(Model)、逍遥散组(Xiaoyaosan)和米非司酮组(Mifepristone),将4T1细胞株接种于各组小鼠腋下,待成瘤后,除Tumor组外其余各组小鼠均进行慢性束缚应激21天,同时Xiaoyaosan组和Mifepristone组小鼠给予相对应的药物灌胃,Tumor组和Model组小鼠灌胃生理盐水。造模结束后,小鼠麻醉断头处死,测量小鼠瘤体重量和体积、内脏指数;采用ELISA方法检测各组小鼠血清肿瘤标志物糖类抗原ca199(Carbohydrate antigen199,CA199)、癌胚抗原(Carcino-embryonic antigen,CEA)、血管内皮生长因子(Vascular endothelial growth factor,VEGF)的含量,血清多巴胺(Dopamine,DA)和皮质酮(Corticosterone,CORT)的含量,以及肿瘤组织转化生长因子β1(Transforming growth factor-β1,TGF-β1)和白介素10(Interleukin 10,IL-10)的含量。采用免疫组化和Western blot方法检测各组小鼠肿瘤组织巨噬细胞极化标志物诱导型一氧化氮合酶(Inducible nitric oxide synthase,iNOS)和精氨酸酶1(Arginase-1,Arg-1)的表达,以及肿瘤组织中细胞外基质金属蛋白酶诱导剂(Extracellular matrix metalloproteinase inducer,EMMPRIN,CD147)及其下游信号分子基质金属蛋白酶2(Matrix metalloproteinases 2),MMP2、基质金属蛋白酶9(MMP9)和VEGF的表达。结果与Tumor组比较,Model组小鼠肿瘤重量和体积,血清CA199、CEA、VEGF、CORT含量,肿瘤TGF-β1和IL-10含量均显著增加;内脏指数和血清DA含量显著减少;肿瘤巨噬细胞M2型极化标志物Arg-1的表达显著增加,M1型极化标志物iNOS的表达显著降低;肿瘤CD147及其下游信号分子MMP2、MMP9和VEGF蛋白表达显著增加,逍遥散和米非司酮均可有效逆转以上改变。结论慢性束缚应激促小鼠乳腺癌进展的机制与肿瘤相关巨噬细胞M2型极化释放TGF-β1增加、激活CD147及其下游相关信号有关,而逍遥散可缓解应激条件下皮质酮增高引起的巨噬细胞M2型极化,减少TGF-β1的生成,抑制CD147及其下游信号,从而抑制慢性应激引起的小鼠乳腺癌进展。

肺腺癌荷瘤鼠碘^(125)粒子植入治疗对肿瘤细胞凋亡及瘤周辐射的影响

目的探讨肺腺癌荷瘤鼠在^(125)I粒子植入治疗后细胞凋亡的情况以及对瘤周辐射的影响。方法肺腺癌荷瘤鼠60只随机分为3组:A组(1枚^(125)I空源对照组)、B组(1枚^(125)I粒子治疗实验组)、C组(2枚^(125)I粒子治疗实验组)。分别在粒子植入后第7、14、21、28天处死瘤鼠,计算抑瘤率、瘤体及瘤周细胞凋亡指数并进行统计学分析。结果B、C组均随着粒子植入的时间延长,抑瘤率和细胞凋亡指数增高(P<0.05);B、C组在相同时间段的抑瘤率、细胞凋亡指数差异均无统计学意义(P>0.05)。A、B、C 3组瘤周细胞的凋亡指数比较,差异无统计学意义(P>0.05)。结论荷瘤鼠^(125)I粒子植入后随着时间的延长,肿瘤细胞凋亡数增多,抑瘤效果较好,并且瘤周正常细胞并未受到辐射损伤。

超声引导复合式冷热消融系统治疗Hepa1-6瘤鼠的疗效及免疫效应

目的探讨超声引导复合式冷热消融系统治疗Hepa1-6瘤鼠的疗效及免疫效应。方法选取健康C57BL/6J小鼠建立肝癌Hepa1-6小鼠移植瘤模型。将40只Hepa1-6荷瘤小鼠随机分为4组:空白对照组、假手术组、外科手术组及复合式冷热消融治疗组,10只/组。分别于治疗前及治疗后1、2、3、4周,采用超声评估各治疗组荷瘤小鼠皮下肿瘤体积变化,采用血流式细胞术检测检查各组小鼠外周血CD4+、CD8+T淋巴细胞数百分比。于治疗2周后随机处死各治疗组小鼠各3只,并对治疗后病灶行HE染色病理检查及免疫组化检测组织CD4+、CD8+细胞光密度值。结果空白对照组及假手术组各时间点的肿瘤组织逐渐增大,但两者差异无统计学意义(P>0.05);与空白对照组及假手术组相比,复合式冷热消融组肿瘤体积在治疗后各时间点肿瘤逐渐缩小,差异有统计学意义(P<0.05)。复合式冷热消融治疗组术后与术前比较,术后各时间点血清CD4+T、CD8+T细胞水平均升高,明显高于空白对照组、假手术组、外科手术组(P<0.05)。冷冻治疗组血清CD4+T、CD8+T细胞水平在治疗后3周出现峰值,随后出现下降。与对照组及假手术组相比,冷冻治疗组在治疗后2周肿瘤组织CD4+、CD8+细胞明显增多,差异有统计学意义(P<0.05)。结论超声引导复合式冷热消融系统治疗Hepa1-6瘤鼠具有很好的治疗疗效,同时还能够激活免疫系统,增强机体的抗肿瘤免疫效应。

STUDY ON GLYCOCONJUGATE CHANGES ON CELL SURFACE IN PROGRESSIVE DEVELOPMENT OF PULMONARY TUMOR

Aim: To investigate glycoconjugate changes on the cell surface of proliferative lesions and neoplasms of mice lungs at various stages of tumorigenesis, the relation between progressive development of mouse pulmonary tumors and expression of cell surface saccharide. Materials and methods: Thirty - one male A/J strain mice at 5 weeks of age were treated intraperitoneally with a single injection of 20 - methylcholanthrene (20 - MC), 292 pulmonary lesions including 31 hyperplasias, 145 alveolar adenomas, 61 papillary adenomas, 55 papillary adenocarcinomas and their combined type were obtained. The binding affinities of cells in normal respiratory epithelia and in proliferative lesions to four peroxidases - conjugated lectins, Maclura pomifera agglutinin (MPA), Arachis hypogea agglutinin (PNA), Ricinus communis agglutinin (RCA), and wheat germ agglutinin (WGA) were examined. Results: Cells of hyperplasia and alveolar adenoma showed fairly strong affinity to all the four lectins. However, part of papillary adenoma cells and greater part of papillary adenocarcinoma cells lost their binding affinity to MPA, PNA, and RCA, but not to WGA. The bindings of MPA, PNA and RNA were detected predominently on the luminal surfaces of benign tumors but not on the luminal surfaces of malignant tumors. WGA might bind to varied types of benign and malignant tumors. Pretreated with neuraminidase, the lesions enhanced the staining intensity for the four lectins, the binding sites of WGA to malignant tumor cells were numerous. A distinct difference in lectin binding affinity between hyperplasia / alveolar adenoma/papillary adenoma and papillary adenocarcinoma was clearly shown( x2 = 46.89, P ＜ 0.01, x2 = 36.77, P ＜ 0.01 and x2 = 52.87, P ＜ 0.01 ) in this experiment. The complex glycoconjugates on the cell surface of malignant and benign lesions during the development of pulmonary tumor were changed,malignant tumor cells differed from the surface of benign tumor cells, the levels of total sialic acid were higher in malignant tumor cells than in benign lesions. Conclusions: The expressions of glycoconjugates on cell surface by lectins may assist differential diagnosis of the malignant and benign lung lesions.

EFFECT OF EPIMEDIUM FLAVONE TOTAL(EFT) ON THE IMMUNOFUNCTION OF EHRLICH ASCITES CANCER(EAC) BEARING MICE

Effect of Epimedium Flavone Total(EFT) on the immunofunction of mice which was burdened with Ehrlich Ascites Cancer (EAC) was studied. The results showed that EFT could increase phagocytosis activity of macrophagocyte (P<0 05), promote lymphocyte transformation rate (P<0 01), increased the sero antibody level meanly (P<0 05), low dosage group could increased Plaque Forming Cell (PFC) signficantly (P<0 05), raise the ratio of spleen to body (P<0 05) of EAC bearing mice. EFT may be enhance burdened mice's immunofunction. Low dosage can improve sero antibody level. Effect on regulating cellular immunofunction are related to dosage. EFT may be inhibit the growth of EAC cells to some degree.

靶向FOLR1的脱氧核酶提高鼻咽癌移植瘤对紫杉醇的敏感性

目的:探讨裸鼠鼻咽癌移植瘤能否通过靶向FOLR1的脱氧核酶(DrzE)提高其对紫杉醇的敏感性。方法:取CNE-1鼻咽癌亲本细胞及CNE-1/taxol紫杉醇耐药细胞进行裸鼠成瘤实验,分别予靶向FOLR1的“10-23”型DrzE单用和紫杉醇单用及紫杉醇-DrzE联用后比较各组鼻咽癌移植瘤体的抑制情况。RT-qPCR及Western blot检测用药前后各组瘤体FOLR1 mRNA及蛋白的表达。结果:CNE-1瘤体较CNE-1/taxol瘤体生长速度快,瘤体体积更大(P<0.05);CNE-1瘤体生长能被紫杉醇单药抑制,但紫杉醇对CNE-1/taxol瘤体无效;两种瘤体生长均能被DrzE单药抑制,其中耐药瘤体更显著(P<0.05);紫杉醇与DrzE联用较分别单用两药对移植瘤的抑制作用更显著(P<0.05);给药前CNE-1/taxol移植瘤中FOLR1的表达量显著高于CNE-1(P<0.01),用药后DrzE单药组及紫杉醇与DrzE联用组中两种移植瘤体FOLR1表达量均显著低于对照组(P<0.05)。结论:靶向FOLR1的脱氧核酶DrzE转染裸鼠移植瘤体后可减慢鼻咽癌瘤体的生长,提高鼻咽癌耐药移植瘤体对紫杉醇的敏感性。