CAJ | 학술논문

目的研究选择性巨噬细胞移动抑制因子(MIF)互变异构酶活性抑制剂ISO-1对BALB/c小鼠大肠癌肝转移的影响及其可能机制. 方法微孔迁移法检测ISO-1对CT26细胞体外侵袭的影响.盲肠造疝原位移植瘤块术建立小鼠大肠癌肝转移模型,将30只成功建模小鼠分为3组,每组10只.每周两次腹腔注射相同体积ISO-1(0.2 ml,20 mg/kg)、5%DMSO和无菌生理盐水(NS)溶液.治疗4周后处死小鼠,肝脏连续病理切片、HE染色,比较各组的肝转移率.L-多巴色素甲酯测定小鼠血清MIF互变异构酶活性;ELISA测定血清中血管内皮生长因子(VEGF)水平;CD31染色标记肿瘤微血管内皮细胞测定肿瘤微血管密度(MVD).结果 100 μmol/L ISO-1作用24 h后CT26细胞穿透聚碳酸酯膜细胞数显著减少[(151±19)比(178±9),P<0.01].ISO组比DMSO组小鼠血清MIF互变异构酶活性为51%比81%,P<0.01.ISO-1组、DMSO组与NS组肝转移率分别为10%、60%与70%(x2=8.30,P<0.05).ISO-1组、DMSO组与NS组小鼠血清VEGF的水平分别为(15±7)pg/ml、(63±10)pg/ml与(67±8)pg/ml,P<0.01.ISO-1组、DMSO组与NS组原位肿瘤组织MVD分别为17±4、36±7与38±5,P<0.01.结论在体外ISO-1减少了CT26细胞的迁移,在体内降低了大肠癌肝转移的发生.其可能机制为ISO-1抑制MIF互变异构酶活性,下调VEGF表达,减少MVD.
목적 연구선택성거서세포이동억제인자(MIF)호변이구매활성억제제ISO-1대BALB/c소서대장암간전이적영향급기가능궤제. 방법미공천이법검측ISO-1대CT26세포체외침습적영향.맹장조산원위이식류괴술건립소서대장암간전이모형,장30지성공건모소서분위3조,매조10지.매주량차복강주사상동체적ISO-1(0.2 ml,20 mg/kg)、5%DMSO화무균생리염수(NS)용액.치료4주후처사소서,간장련속병리절편、HE염색,비교각조적간전이솔.L-다파색소갑지측정소서혈청MIF호변이구매활성;ELISA측정혈청중혈관내피생장인자(VEGF)수평;CD31염색표기종류미혈관내피세포측정종류미혈관밀도(MVD).결과 100 μmol/L ISO-1작용24 h후CT26세포천투취탄산지막세포수현저감소[(151±19)비(178±9),P<0.01].ISO조비DMSO조소서혈청MIF호변이구매활성위51%비81%,P<0.01.ISO-1조、DMSO조여NS조간전이솔분별위10%、60%여70%(x2=8.30,P<0.05).ISO-1조、DMSO조여NS조소서혈청VEGF적수평분별위(15±7)pg/ml、(63±10)pg/ml여(67±8)pg/ml,P<0.01.ISO-1조、DMSO조여NS조원위종류조직MVD분별위17±4、36±7여38±5,P<0.01.결론 재체외ISO-1감소료CT26세포적천이,재체내강저료대장암간전이적발생.기가능궤제위ISO-1억제MIF호변이구매활성,하조VEGF표체,감소MVD.
Objective To investigate the effects of ISO-1, a selective MIF tautomerase activity inhibitor, on liver metastasis in a BALB/c mouse model of colonic cancer. Methods Micmporous migration assay was used to determine the effect of ISO-1 on the invasion abilities of CT26 cells. Orthotopic transplantation of fresh colonic tumor fragments into the hernial sac of cecum was used in a BALB/c mouse model of eolorectal cancer. Thirty mouse models were divided into three groups and treated respectively with ISO-1 (0. 2 ml, 20 mg/kg), 5% DMSO and NS ( normal sodium) twice a week, iutraperitoneally. After 4 weeks, mice were sacrificed and the whole livers were made into serial slices to detect the occurrence of liver metastasis. Serum MIF tautomerase activities were measured using L-dopachrome methyl ester, ELISA was used to test serum VEGF concentrations. Immunohistochemical staining of CD31 was used for comparing microvascular density (MVD) of tumor tissues. Results 100 μmol/L ISO-1 treatment for 24 hours significantly reduced the average number of the cells penetrating polycarbonates, ( 151 ± 19 ) vs. ( 178 ± 9 ), P<0. 01. Serum MIF tautomerase activities were significantly inhibited after ISO-1 treatment (51% vs. 81%, P <0. 01 ). Compared with DMSO and NS treatment, ISO-1 decreased the occurrence of liver metastasis ( 10% ,60% and 70% ,respectively;x2 = 8. 30, P < 0. 05 ). Also ISO-1 decreased serum VEGF levels ( 15 ± 7 ) pg/ml, ( 63 ± 11 ) pg/ml and ( 67 ± 8 ) pg/ml, respectively; P < 0. 01 and the MVD of tumor tissues (17±4) ,(36±7) and( 38±5) ,respectively; P<0. 01. Conclusion In vitro ISO-1 inhibits the invasion ability of CT26 cells. In vivo ISO-1 reduces the occurrence of liver metastasis, possibly by a mechanism of inhibiting MIF tautomerase activities, down-regulating the expression of VEGF and reducing MVD.