CAJ | 학술논문

目的探讨转染CD40配体(CIMOL)的卵巢癌细胞株OVHM(CD40L-OVHM)体内抗卵巢癌肝转移的可能机制.方法 6～8周龄的C57BL/6N×C3H/He杂交一代(B6C3F1)雌性小鼠5只,经小鼠脾脏内接种OVHM,应用HE染色法验证小鼠肝脾转移模型是否建立成功.将OVHM细胞、空载体DNA-pMKITneo-OVHM细胞和CD40L-OVHM细胞接种于B6C3F1小鼠的脾脏,应用流式细胞术,分析荷瘤小鼠脾细胞CD11c分子的表达情况;应用四甲基偶氮唑蓝(MTT)法,检测荷瘤小鼠脾脏细胞毒性T淋巴细胞(CTL)的杀伤活性;应用酶联免疫吸附试验(ELISA),检测荷瘤小鼠外周血血清中干扰素(IFN)-γ、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-12、IL-4和IL-10的含量,并且观察小鼠脾脏和肝脏的成瘤情况及小鼠生存时间.结果经病理学证实,卵巢癌肝脾转移动物模型建立成功.CD40L-OVHM组中小鼠脾细胞CD11c阳性细胞率明显高于OVHM组和转染空载体DNA-pMKITneo-OVHM组.CD40L-OVHM组小鼠脾脏内CTL的特异性杀伤活性明显增强,小鼠外周血血清中IFN-γ、TNF-α和IL-12的含量明显升高,而IL-4和IL-10的含量则明显降低,与OVHM组和转染空载体DNA-pMKITneo-OVHM组相比,差异均有统计学意义(P<0.05).CD40L-OVHM组小鼠的肝脏和脾脏重量均明显低于OVHM组和转染空载体DNA-pMKITneo-OVHM组,并且小鼠的生存期也明显延长(P<0.05).结论转染CD40L cDNA的卵巢癌细胞可促进脾脏树突状细胞(DC)的成熟和分化,增强脾脏CTL的特异性杀伤活性,诱导Th1型细胞因子的分泌,抑制Th2型细胞因子的分泌,这可能是CD40L基因体内抗卵巢癌肝转移的机制之一.
목적 탐토전염CD40배체(CIMOL)적란소암세포주OVHM(CD40L-OVHM)체내항란소암간전이적가능궤제.방법 6～8주령적C57BL/6N×C3H/He잡교일대(B6C3F1)자성소서5지,경소서비장내접충OVHM,응용HE염색법험증소서간비전이모형시부건립성공.장OVHM세포、공재체DNA-pMKITneo-OVHM세포화CD40L-OVHM세포접충우B6C3F1소서적비장,응용류식세포술,분석하류소서비세포CD11c분자적표체정황;응용사갑기우담서람(MTT)법,검측하류소서비장세포독성T림파세포(CTL)적살상활성;응용매련면역흡부시험(ELISA),검측하류소서외주혈혈청중간우소(IFN)-γ、종류배사인자(TNF)-α、백세포개소(IL)-12、IL-4화IL-10적함량,병차관찰소서비장화간장적성류정황급소서생존시간.결과 경병이학증실,란소암간비전이동물모형건립성공.CD40L-OVHM조중소서비세포CD11c양성세포솔명현고우OVHM조화전염공재체DNA-pMKITneo-OVHM조.CD40L-OVHM조소서비장내CTL적특이성살상활성명현증강,소서외주혈혈청중IFN-γ、TNF-α화IL-12적함량명현승고,이IL-4화IL-10적함량칙명현강저,여OVHM조화전염공재체DNA-pMKITneo-OVHM조상비,차이균유통계학의의(P<0.05).CD40L-OVHM조소서적간장화비장중량균명현저우OVHM조화전염공재체DNA-pMKITneo-OVHM조,병차소서적생존기야명현연장(P<0.05).결론 전염CD40L cDNA적란소암세포가촉진비장수돌상세포(DC)적성숙화분화,증강비장CTL적특이성살상활성,유도Th1형세포인자적분비,억제Th2형세포인자적분비,저가능시CD40L기인체내항란소암간전이적궤제지일.
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.