CAJ | 학술논문

目的探讨靶向巨噬细胞移动抑制因子(MIF)的小干扰RNA (siRNA)对BALB/C小鼠大肠癌肝转移的影响及其可能的机制。方法盲肠造疝原位接种瘤块法建立BALB/C小鼠大肠癌肝转移模型。将成功建模的小鼠随机分为3组。分别每周2次肿瘤原位注射相同体积的靶向MIF的siRNA( MIF siRNA,0.15 ng/kg)、非特异性siRNA( NS-siRNA,0.15 ng/kg)及生理盐水(NS,0.15 ng/kg),治疗4周。治疗结束3d后,处死小鼠。肝脏连续切片,苏木素-伊红(HE)染色观察各组大肠癌肝转移;酶联免疫吸附试验(ELISA)检测血清MIF及血管内皮生长因子(VEGF)浓度;免疫组织化学染色检测肝转移灶微血管密度(MVD)。结果 MIF siRNA组、NS-siRNAR组及NS组的大肠癌肝转移率分别为10％、60％与70％(x2=8.30,P＜0.05),小鼠血清MIF分别为(20±4)、(72±8)与(78 ±7) ng/L(P ＜0.05);小鼠血清VEGF分别为(20±4)、(77±9)与(77±10) ng/L(P＜0.05);肝转移灶的MVD分别为19±3、29 ±6与35±7(P＜0.05)。结论靶向MIF的siRNA降低了小鼠大肠癌肝转移的发生率,其机制可能是靶向MIF的siRNA抑制了MIF表达,下调VEGF的表达,减少了MVD。
목적 탐토파향거서세포이동억제인자(MIF)적소간우RNA (siRNA)대BALB/C소서대장암간전이적영향급기가능적궤제。방법 맹장조산원위접충류괴법건립BALB/C소서대장암간전이모형。장성공건모적소서수궤분위3조。분별매주2차종류원위주사상동체적적파향MIF적siRNA( MIF siRNA,0.15 ng/kg)、비특이성siRNA( NS-siRNA,0.15 ng/kg)급생리염수(NS,0.15 ng/kg),치료4주。치료결속3d후,처사소서。간장련속절편,소목소-이홍(HE)염색관찰각조대장암간전이;매련면역흡부시험(ELISA)검측혈청MIF급혈관내피생장인자(VEGF)농도;면역조직화학염색검측간전이조미혈관밀도(MVD)。결과 MIF siRNA조、NS-siRNAR조급NS조적대장암간전이솔분별위10％、60％여70％(x2=8.30,P＜0.05),소서혈청MIF분별위(20±4)、(72±8)여(78 ±7) ng/L(P ＜0.05);소서혈청VEGF분별위(20±4)、(77±9)여(77±10) ng/L(P＜0.05);간전이조적MVD분별위19±3、29 ±6여35±7(P＜0.05)。결론 파향MIF적siRNA강저료소서대장암간전이적발생솔,기궤제가능시파향MIF적siRNA억제료MIF표체,하조VEGF적표체,감소료MVD。
Objective To investigate the effects of macrophage migration inhibitory tactor( MIF)small interfering RNA (siRNA), a siRNA targeting MIF on hepatic metastasis in a BALB/C mouse model of colonic cancer. Methods Orthotopic transplantation of fresh colorectal cancer tumor fragments into hernial sac of cecum was used in a BALB/C mouse model of colorectal cancer. Thirty mouse models were divided randomly into three groups and treated respectively with MIF siRNA (0. 15 ng/kg), NC-MIF siRNA and normal saline (NS) twice a week, intratumorally. After 4th week, mice were sacrificed and the total livers were made into serial slices to detect the occurrence of hepatic metastasis. Enzyme linked immunosorbent assay (ELISA) was used to test the serum MIF and vascular endothelial growth factor (VEGF) concentrations. Immunohistochemical staining of CD31 was used for comparing microvessel density of the tumor tissue. Results Compared with NC-MIF siRNA and NS groups, MIF siRNA decreased the occurrence of hepatic metastasis ( 10％, 60％ and 70％, respectively ;x2 =8. 30 ,P ＜ 0. 05 ). Also MIF siRNA decreased serum MIF levels[(20 ±4), (72 ±8) and (78 ±7) ng/L, respectively,P ＜0. 05]and VEGF levels[(20 ± 4), (77 ± 9), (77 ± 10) ng/L, respectively, P ＜ 0. 05]. The MVD of hepatic tumor tissues in MIF-siRNA, NC-MIF siRNA and NS groups was 19 ± 3, 29 ± 6 and 35 ± 7, respectively (P ＜ 0. 05).Conclusion MIF siRNA reduced the occurrence of hepatic metastasis, possibly by a mechanism of MIF siRNA inhibiting the expression of MIF, down-regulating the expression of VEGF and reducing MVD.