CD40-CD40L与肿瘤

CD40和其配体CD40L(又称CD154)是体内特异性免疫反应系统重要的一对共刺激分子,分别属于TNF受体和TNF超家族成员。CD40／CD40L参与细胞免疫及体液免疫,在炎症反应,自身免疫性疾病,心血管疾病中起重要作用。不仅如此,CD40还表达于多种肿瘤细胞表面,可利用CD40L与其结合用于肿瘤治疗。我们就CD40-CD40L在肿瘤中的应用进行了综述。     

CD40-CD40L与肿瘤

CD40和其配体CD40L(又称CD154)是体内特异性免疫反应系统重要的一对共刺激分子,分别属于TNF受体和TNF超家族成员.CD40/CD40L参与细胞免疫及体液免疫,在炎症反应,自身免疫性疾病,心血管疾病中起重要作用.不仅如此,CD40还表达于多种肿瘤细胞表面,可利用CD40L与其结合用于肿瘤治疗.我们就CD40-CD40L在肿瘤中的应用进行了综述.     

CD40配体激活抑制肺癌细胞生长机制的实验研究

目的探讨CD40配体激活对肺癌细胞生长、增殖的影响。方法采用CCK-8法检测不同浓度sCD40L作用后A549肺癌细胞株存活率,选定最佳干预浓度;流式细胞术检测sCD40L最佳干预浓度作用后肺癌细胞株A549、SPC-A-1细胞周期变化。结果sCD40L的最佳干预浓度为2μg/ml,与肺癌细胞株A549、SPC-A-1共培养48 h,sCD40L可使高表达CD40的A549细胞生长抑制,细胞周期阻滞在G1期,而对CD40低表达的SPC-A-1细胞生长无明显影响。结论sCD40L通过阻滞细胞周期可明显抑制CD40高表达肺癌细胞的生长。     

重组腺病毒鼠CD40L对小鼠淋巴瘤治疗和预防作用的体内研究

目的:探讨重组腺病毒鼠CD40-ligand基因(Adv-mCD40L)对小鼠淋巴瘤的免疫治疗作用及免疫预防作用。方法:构建含有小鼠CD40L基因的重组腺病毒载体,体外转染小鼠B淋巴瘤细胞(A20细胞)。流式细胞术检测A20细胞转染前后表面CD80、CD86、CD40L(CD154)、CD40表达情况。在体内实验中,治疗组首先建立小鼠A20荷瘤模型,分别于肿瘤内多点注射Adv-mCD40L和Adv;预防组小鼠预先分别接种负载A20细胞抗原的Adv-mCD40L和Adv,免疫后再次接种具有活性的A20细胞,均以生理盐水作为对照,观察小鼠成瘤时间及肿瘤生长情况。结果:体外转染mCD40L基因24h内CD154和CD40表达水平改变,48h后A20表达CD80、CD86增强。经Adv-mCD40L免疫后的小鼠成瘤时间较其他组明显延迟。经mCD40L基因治疗的小鼠均可见瘤体生长速度明显受抑,肿瘤大小分别与经Adv及NS治疗的小鼠比较,有显著差异(P<0.05)。结论:转染mCD40L基因可以增强A20表面协同刺激分子的表达,增强A20细胞的免疫原性,激活机体的抗肿瘤免疫,延缓成瘤时间,抑制肿瘤生长。     

Role for CD40L in the Therapy of Human Cancer

CD40L-CD40 interaction is central to the control of thymusdependent humoral immunity and cell mediated immune responses. CD40, a member of the tumor necrosis factor receptor （TNF- R） family, has been found on the surface of B lymphocytes, monocytes, hematopoietic progenitors, dendritic cells （DCs）, endothelial cells, epithelial cells and so on. Its natural ligand （CD40 ligand, CD40L）, CD154, a member of the tumor necrosis factor （TNF） family, is mainly expressed on activated CD4^＋ T lymphocytes. A direct growth-inhibitory effect can be found when ligated CD40 is on human breast, ovarian, cervical, bladder, non-small cell lung, and squamous epithelial carcinoma cells. This effect is related to the induction of cell cycle blockage and/or apoptosis. CD40L induces phenotypic and functional maturation of DCs, and can increase tumor immunogenicity through up-regulation of costimulatory molecular expression and cytokine production by epithelial cancer cells. As a result, CD40L can enhance tumor rejection immune responses. Furthermore, by means of a “bystander effect”, even CD40-negative tumor subsets can be eliminated by activated tumor-reactive cytotoxic T lymphocytes（CTL）. This review summarizes recent findings on CD40L recombinant protein and gene therapy-based tumor treatment approaches.     

结肠癌组织中CD40蛋白表达及其生物学意义

目的:研究免疫共刺激分子CD40在结肠癌及癌旁组织中的表达情况及其生物学意义. 方法:应用免疫组织化学技术检测65例结肠癌组织和10例癌旁组织中CD40蛋白的表达,并分析CD40表达水平与临床病理特征的相关性.结果: 免疫组织化学检测结果显示,CD40蛋白阳性染色于细胞膜和细胞质,在结肠癌组织中高表达率为41.5%,而在正常癌旁组织中不表达,差异有统计学意义(P<0.05).CD40蛋白表达与肿瘤TNM分级和淋巴结转移显著相关(P<0.05),与患者年龄、性别及肿瘤大小无相关性(P>0.05).结论:CD40分子可能在结肠癌发病过程中起重要作用,可作为结肠癌预后判断及淋巴结转移监测的有价值的指标之一.     

B-NHL患者骨髓单个核细胞表面CD40、CD40L 的表达及意义

目的探讨B细胞非霍奇金淋巴瘤（BcellNon—Hodgkin’SLymphoma,B—NHL）患者骨髓单个核细胞（BMMC）表面CIMO、CIM-0L的表达情况及变化规律,分析CIM0、CIMOI。表达与淋巴瘤发病、侵袭性及预后的关系及意义。方法采集20例初诊B—NHL伴有骨髓侵犯的患者（实验组）的骨髓标本及12例健康者（对照组）的骨髓标本,分离单个核细胞,采用直接荧光抗体标记,应用流式细胞仪检测细胞表面C1M0、CIMOL的表达情况。相关实验结果进行统计学比较。结果（1）B—NHL中CIM0阳性率（45．78±3．82）％显著高于正常对照组（13．99±2．20）％,P〈0．01;B—NHL中CIMOL阳性率（3．44±0．88）％显著低于正常对照组（10．64±1．03）％,P〈0．01;（2）B—NHL伴有骨髓侵犯的患者CIM0、CIMOL阳性率与病理类型、LDH值、全身症状呈显著相关,P〈0．05;（3）B—NHL伴有骨髓侵犯的患者中CIMO与CIMOL表达呈负相关,r=-0．47,P〈0．05。结论（1）CIM0、CIM-0L阳性率与病理类型、LDH值、全身症状密切相关,其可能与B—NHL的侵袭性及预后有关;（2）B—NHL患者中CIM0表达的增高、CIMOL表达的减少可能是影响其发病的因素之一;（3）B—NHL患者中CD40L表达的减少可能与肿瘤免疫逃逸机制有关。     

CD40单克隆抗体的抗肿瘤机制及其应用

CD40是分子量为45—50KD的Ⅰ型跨膜糖蛋白,属于肿瘤坏死因子受体（TNFR）超家族成员,表达于多种抗原递呈细胞、内皮细胞、上皮细胞、造血前体细胞、成纤维细胞以及某些肿瘤细胞。通过单克隆抗体激发CD40分子不仅可以直接作用于CD40^＋分子的肿瘤细胞,其介导的信号还可在多个环节影响肿瘤发生、发展,如调控细胞增殖／凋亡,增强其它治疗手段的敏感性,并能通过调节适应性和固有免疫应答而起到抗肿瘤作用,广泛应用于肿瘤治疗。     

可溶性CD40配体对肺癌细胞 A549的生物学作用及其机制研究

背景与目的:尽管对 CD40分子在 B细胞中的功能已有深入的研究,但 CD40在肺癌细胞中的功能目前知之甚少.本研究旨在探讨可溶性 CD40配体(solublE-CD40 ligand, sCD40L)对肺癌细胞株 A549(CD40表达阳性细胞株)的生物学作用及其相关机制.方法:采用四氮唑盐(MTT)比色、 3H标记胸腺脱氧嘧啶核苷( 3H-TdR)掺入法检测 sCD40L对 A549细胞增殖的影响,免疫荧光标记和流式细胞术测定细胞表型及细胞周期的改变,流式细胞术、逆转录聚合酶链式反应( RT-PCR)及 Western blot分析测定 sCD40L对 A549细胞凋亡的影响及 Bcl-2、 Bax基因表达的变化.结果:(1) sCD40L可抑制 A549细胞的增殖(与对照组比较 P< 0.05).(2) sCD40L作用 72 h后, A549细胞表面 CD49e、 CD54、 TNFRⅠ及 CD95L的表达 [分别为( 61.2± 4.8)%,( 31.2± 6.1)%,( 42.7± 5.9)%,( 38.2± 3.4)% ]较对照组 [分别为 (34.7± 2.1)%, (7.1± 1.6)%, (15.2± 4.1)%, (10.1± 2.3)% ]明显升高,而 TNFRⅡ的表达 [( 8.7± 0.8)% ]较对照组 [( 58.1± 3.6)% ]下降.( 3) sCD40L作用 72h后, A549细胞 G1 期细胞比例 [(76.0± 9.1)% ]较对照组 [(56.7± 6.9)% ]增加, S期细胞比例 [(10.3± 5.7)% ]较对照组 [(32.7± 5.5)% ]减少. ( 4) sCD40L在短期( 72 h)内并不引起 A549细胞明显凋亡,但可上调 Bax的表达.结论: sCD40L可引起肺癌细胞 A549生长抑制,细胞周期和表型改变,并可引起凋亡相关基因表达的改变.     

胃癌中MCP-1和CD40L的表达及联合作用对PGE2及VEGF产物的生物学影响

目的：探讨在胃癌间质细胞中单核细胞趋化蛋白（MCP-1）和CD40L配体（CD40L）的表达及它们之间的相互作用,以及对前列腺素E2（PGE2）及血管内皮生长因子（VEGF）产物的影响。方法：应用RT—PCR技术测定胃癌组织中CD40L和MCP-1的mRNA水平。在巨噬细胞中,分别用MCP-1、CD40L、MCP-1和CD40L共同刺激后,测定COX-2、PGE2和VEGF产物。结果：在MCP-1和CD40L共同刺激的巨噬细胞中,COX-2表达水平显著升高,同时与PGE2和VEGF产物升高相互关联。在COX-2表达上,MCP-1和CD40L巨噬细胞有协同作用,随之PGE2和VEGF产物也增加。与幽门螺杆菌感染的胃炎患者相比,低分化胃癌患者中CD40L和MCP-1的mRNA表达水平明显升高。结论：胃癌中MCP-1和CD40L在COX-2表达上有协同作用,因此对PGE2及VEGF产物也有协同作用。     

表达CD40L基因的卵巢癌细胞诱导树突状细胞成熟及分泌Th1型细胞因子的作用机制

目的 探讨转染CD40L cDNA的卵巢癌细胞株OVHM对树突状细胞(DC)成熟、分化的影响及诱导其分泌细胞因子的机制.方法 采用脂质体转染法将鼠全长CD40L基因转染人小鼠卵巢癌细胞株OVHM中,用G418筛选出表达CD40L基因的抗药性克隆细胞(CD40L-OVHM).应用免疫磁珠分选并纯化小鼠骨髓DC.将DC与转染和未转染CD40L cDNA的OVHM细胞混合培养,流式细胞术检测DC细胞表面人主要组织相容性复合物Ⅰ类分子(MHC-Ⅰ)、Ⅱ类分子(MHC-Ⅱ)、CD80、CD86和CCR7的表达,逆转录聚合酶链反应(RT-PCR)检测共培养细胞中白细胞介素(IL)-12、IL-23、IL-27、IL-18、γ干扰素(IFN-γ)、Mig基因的表达.结果 DC与CD40L-OVHM细胞混合培养后可形成集落,且上调了DC细胞MHC-Ⅰ、MHC-Ⅱ、CD80、CD86、CCR7的共刺激分子和黏附分子的表达.在DC+CD40L-OVHM组可检测到细胞因子IL-12、IL-23、IL-27、IL-18、IFN-γ、Mig基因的表达,而在DC组、DC+OVHM组、CD40L-OVHM组、OVHM组未检测到这些基因的表达.结论 表达CD40L的卵巢癌细胞促进了DC的成熟,诱导了Th1型细胞因子的分泌,这可能是转染CD40L基因的卵巢癌细胞产生抗肿瘤效应的机制之一.     

转染CD40配体的卵巢癌细胞株OVHM抗卵巢癌肝转移机制的研究

Objective To examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murinc ovarian cancer OVUM cells (CD4OL-OVHM) injected into the spleen on liver metastasis in mice. Methods OVUM cells were inoculated into the spleen of 6 ～ 8 week-old female B6C3F1 (C57BL/6N × C3H/He) mice. The established liver metastasis was identified by histopathology (HE staining). OVUM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CD11c in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-γ, TNF-α, IL-12, IL4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded. Results The mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11c and the specific killing rote in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-γ, TNF-α and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo-OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly (P < 0.05). The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group (P < 0.05). Conclusion The data directly demonstrate that the expression of CD40L in ovarian cancer cells (CD40L-OVHM) can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune function of peripheral blood cells and the immune balance between Thl cells and Th2 cells, which maybe the possible mechanism induced by CD40L in mice inhibiting the development of liver metastasis.     

转染CD40配体的卵巢癌细胞株OVHM抗卵巢癌肝转移机制的研究

Objective To examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murinc ovarian cancer OVUM cells (CD4OL-OVHM) injected into the spleen on liver metastasis in mice. Methods OVUM cells were inoculated into the spleen of 6 ～ 8 week-old female B6C3F1 (C57BL/6N × C3H/He) mice. The established liver metastasis was identified by histopathology (HE staining). OVUM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CD11c in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-γ, TNF-α, IL-12, IL4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded. Results The mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11c and the specific killing rote in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-γ, TNF-α and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo-OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly (P < 0.05). The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group (P < 0.05). Conclusion The data directly demonstrate that the expression of CD40L in ovarian cancer cells (CD40L-OVHM) can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune function of peripheral blood cells and the immune balance between Thl cells and Th2 cells, which maybe the possible mechanism induced by CD40L in mice inhibiting the development of liver metastasis.     

转染CD40配体的卵巢癌细胞株OVHM抗卵巢癌肝转移机制的研究

Objective To examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murinc ovarian cancer OVUM cells (CD4OL-OVHM) injected into the spleen on liver metastasis in mice. Methods OVUM cells were inoculated into the spleen of 6 ～ 8 week-old female B6C3F1 (C57BL/6N × C3H/He) mice. The established liver metastasis was identified by histopathology (HE staining). OVUM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CD11c in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-γ, TNF-α, IL-12, IL4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded. Results The mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11c and the specific killing rote in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-γ, TNF-α and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo-OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly (P < 0.05). The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group (P < 0.05). Conclusion The data directly demonstrate that the expression of CD40L in ovarian cancer cells (CD40L-OVHM) can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune function of peripheral blood cells and the immune balance between Thl cells and Th2 cells, which maybe the possible mechanism induced by CD40L in mice inhibiting the development of liver metastasis.     

转染CD40配体的卵巢癌细胞株OVHM抗卵巢癌肝转移机制的研究

Objective To examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murinc ovarian cancer OVUM cells (CD4OL-OVHM) injected into the spleen on liver metastasis in mice. Methods OVUM cells were inoculated into the spleen of 6 ～ 8 week-old female B6C3F1 (C57BL/6N × C3H/He) mice. The established liver metastasis was identified by histopathology (HE staining). OVUM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CD11c in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-γ, TNF-α, IL-12, IL4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded. Results The mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11c and the specific killing rote in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-γ, TNF-α and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo-OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly (P < 0.05). The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group (P < 0.05). Conclusion The data directly demonstrate that the expression of CD40L in ovarian cancer cells (CD40L-OVHM) can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune function of peripheral blood cells and the immune balance between Thl cells and Th2 cells, which maybe the possible mechanism induced by CD40L in mice inhibiting the development of liver metastasis.     

转染CD40配体的卵巢癌细胞株OVHM抗卵巢癌肝转移机制的研究

Objective To examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murinc ovarian cancer OVUM cells (CD4OL-OVHM) injected into the spleen on liver metastasis in mice. Methods OVUM cells were inoculated into the spleen of 6 ～ 8 week-old female B6C3F1 (C57BL/6N × C3H/He) mice. The established liver metastasis was identified by histopathology (HE staining). OVUM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CD11c in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-γ, TNF-α, IL-12, IL4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded. Results The mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11c and the specific killing rote in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-γ, TNF-α and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo-OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly (P < 0.05). The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group (P < 0.05). Conclusion The data directly demonstrate that the expression of CD40L in ovarian cancer cells (CD40L-OVHM) can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune function of peripheral blood cells and the immune balance between Thl cells and Th2 cells, which maybe the possible mechanism induced by CD40L in mice inhibiting the development of liver metastasis.     

转染CD40配体的卵巢癌细胞株OVHM抗卵巢癌肝转移机制的研究

Objective To examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murinc ovarian cancer OVUM cells (CD4OL-OVHM) injected into the spleen on liver metastasis in mice. Methods OVUM cells were inoculated into the spleen of 6 ～ 8 week-old female B6C3F1 (C57BL/6N × C3H/He) mice. The established liver metastasis was identified by histopathology (HE staining). OVUM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CD11c in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-γ, TNF-α, IL-12, IL4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded. Results The mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11c and the specific killing rote in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-γ, TNF-α and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo-OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly (P < 0.05). The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group (P < 0.05). Conclusion The data directly demonstrate that the expression of CD40L in ovarian cancer cells (CD40L-OVHM) can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune function of peripheral blood cells and the immune balance between Thl cells and Th2 cells, which maybe the possible mechanism induced by CD40L in mice inhibiting the development of liver metastasis.     

转染CD40配体的卵巢癌细胞株OVHM抗卵巢癌肝转移机制的研究

Objective To examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murinc ovarian cancer OVUM cells (CD4OL-OVHM) injected into the spleen on liver metastasis in mice. Methods OVUM cells were inoculated into the spleen of 6 ～ 8 week-old female B6C3F1 (C57BL/6N × C3H/He) mice. The established liver metastasis was identified by histopathology (HE staining). OVUM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CD11c in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-γ, TNF-α, IL-12, IL4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded. Results The mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11c and the specific killing rote in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-γ, TNF-α and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo-OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly (P < 0.05). The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group (P < 0.05). Conclusion The data directly demonstrate that the expression of CD40L in ovarian cancer cells (CD40L-OVHM) can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune function of peripheral blood cells and the immune balance between Thl cells and Th2 cells, which maybe the possible mechanism induced by CD40L in mice inhibiting the development of liver metastasis.     

转染CD40配体的卵巢癌细胞株OVHM抗卵巢癌肝转移机制的研究

Objective To examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murinc ovarian cancer OVUM cells (CD4OL-OVHM) injected into the spleen on liver metastasis in mice. Methods OVUM cells were inoculated into the spleen of 6 ～ 8 week-old female B6C3F1 (C57BL/6N × C3H/He) mice. The established liver metastasis was identified by histopathology (HE staining). OVUM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CD11c in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-γ, TNF-α, IL-12, IL4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded. Results The mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11c and the specific killing rote in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-γ, TNF-α and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo-OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly (P < 0.05). The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group (P < 0.05). Conclusion The data directly demonstrate that the expression of CD40L in ovarian cancer cells (CD40L-OVHM) can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune function of peripheral blood cells and the immune balance between Thl cells and Th2 cells, which maybe the possible mechanism induced by CD40L in mice inhibiting the development of liver metastasis.     

转染CD40配体的卵巢癌细胞株OVHM抗卵巢癌肝转移机制的研究

Objective To examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murinc ovarian cancer OVUM cells (CD4OL-OVHM) injected into the spleen on liver metastasis in mice. Methods OVUM cells were inoculated into the spleen of 6 ～ 8 week-old female B6C3F1 (C57BL/6N × C3H/He) mice. The established liver metastasis was identified by histopathology (HE staining). OVUM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CD11c in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-γ, TNF-α, IL-12, IL4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded. Results The mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11c and the specific killing rote in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-γ, TNF-α and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo-OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly (P < 0.05). The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group (P < 0.05). Conclusion The data directly demonstrate that the expression of CD40L in ovarian cancer cells (CD40L-OVHM) can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune function of peripheral blood cells and the immune balance between Thl cells and Th2 cells, which maybe the possible mechanism induced by CD40L in mice inhibiting the development of liver metastasis.