CAJ | 학술논문

目的研究生长抑素类似物奥曲肽(OCT)与紫杉醇(FTX)的偶合物对人肺腺癌细胞株A549裸鼠移植瘤的靶向治疗作用.方法构建1分子紫杉醇-1分子奥曲肽偶合物(PTX-OCT)和2分子紫杉醇-1分子奥曲肽偶合物(2PTX-OCT),建立裸鼠腋部皮下移植瘤模型,将荷瘤裸鼠随机分为8组.在l、7、21 d尾静脉分别注射150 nmol/kg PTX-OCT;2FTX-OCT;OCT、PTX混合物(OP);OCT;PTX;300 nmol/kg PTX(2PTX)和2PTX-OCT[2(2PTX-OCT)].测各组小鼠体重及肿瘤体积;计算肿瘤倍增时间;眼内眦静脉取血测每组小鼠白细胞数量;组织切片HE染色及核仁区银染(AgNOR)观察肿瘤细胞生长情况;DNA梯度法检测细胞凋亡;RT-PCR法测肿瘤细胞生长抑素受体(SSTR)1至5亚型(SSTRI-SSTR5)mRNA表达情况;免疫组化SP法检测肿瘤微血管密度(MVD)以及生长抑素受体2和5亚型表达情况.结果 2PTX-OCT与2(2PTX-OCT)处理组在3次给药后与对照组相比:肿瘤生长显著抑制(均P<0.01);肿瘤倍增时间延长(均P<0.01).PTX与2PTX-OCT处理组相比,在每次给药后5 d裸鼠体重减轻差异有统计学意义(P<0.05);PTX与2PTX组的白细胞总数在26 d降到最低,与对照组、2PTX-OCT组及2(2VTX-OCT)组相比差异均有统计学意义(均P<0.05).2PTX.OCT组及2(2PTX-OCT)组MVD均明显少于对照组(均P<O.01).DNA梯度法显示2PTX-OCT处理组有明显的DNA片段化的梯状条带.生长抑素受体l、2.4、5亚型的mRNA在荷瘤小鼠体内表达;免疫组化显示SSTR2主要表达于肿瘤细胞的细胞膜上,SSTR5表达于肿瘤细胞的细胞膜和胞质中.结论 2PTX-OCT可显著抑制肿瘤增殖,减少肿瘤微血管形成,显示出良好的治疗作用,有望成为晚期非小细胞肺癌治疗的有效手段之一.
목적 연구생장억소유사물오곡태(OCT)여자삼순(FTX)적우합물대인폐선암세포주A549라서이식류적파향치료작용.방법 구건1분자자삼순-1분자오곡태우합물(PTX-OCT)화2분자자삼순-1분자오곡태우합물(2PTX-OCT),건립라서액부피하이식류모형,장하류라서수궤분위8조.재l、7、21 d미정맥분별주사150 nmol/kg PTX-OCT;2FTX-OCT;OCT、PTX혼합물(OP);OCT;PTX;300 nmol/kg PTX(2PTX)화2PTX-OCT[2(2PTX-OCT)].측각조소서체중급종류체적;계산종류배증시간;안내자정맥취혈측매조소서백세포수량;조직절편HE염색급핵인구은염(AgNOR)관찰종류세포생장정황;DNA제도법검측세포조망;RT-PCR법측종류세포생장억소수체(SSTR)1지5아형(SSTRI-SSTR5)mRNA표체정황;면역조화SP법검측종류미혈관밀도(MVD)이급생장억소수체2화5아형표체정황.결과 2PTX-OCT여2(2PTX-OCT)처리조재3차급약후여대조조상비:종류생장현저억제(균P<0.01);종류배증시간연장(균P<0.01).PTX여2PTX-OCT처리조상비,재매차급약후5 d라서체중감경차이유통계학의의(P<0.05);PTX여2PTX조적백세포총수재26 d강도최저,여대조조、2PTX-OCT조급2(2VTX-OCT)조상비차이균유통계학의의(균P<0.05).2PTX.OCT조급2(2PTX-OCT)조MVD균명현소우대조조(균P<O.01).DNA제도법현시2PTX-OCT처리조유명현적DNA편단화적제상조대.생장억소수체l、2.4、5아형적mRNA재하류소서체내표체;면역조화현시SSTR2주요표체우종류세포적세포막상,SSTR5표체우종류세포적세포막화포질중.결론 2PTX-OCT가현저억제종류증식,감소종류미혈관형성,현시출량호적치료작용,유망성위만기비소세포폐암치료적유효수단지일.
Objective To evaluate the antitumor effects of the conjugates synthesized by coupling cytotoxlc paclitaxel (PTX) to somatostatin analog octreotide (OCT) on human non small cell lung cancer (NSCLC). Methods Two cytotoxic somatostatin analog, PTX-OCT and 2PTX-OCT, were developed in which PIX was linked to octreotide. Forty-five BALB/c nu/nu nude mice were injected with human NSCLC cells of the line A549 into the fight armpit The nude mice that were xenografted were randomly divided into 8 groups. ①control group (n=6), ②FIX-OCT group (n=5), injected intravenously with PTC-OCT 150 nmol/kg on days 1,7, and 21, ③ 2PTX-OCT group (n=6), injected intravenously with PTrc-ocT 150 nmoL/kg, ④ OP group (n=6), injected with mixture of FIX and OCT 150 nmol/kg, ⑤ OCT group (n=5) injected with OCT 150 nmoL/kg ⑤ PTX group (n=6), injected with PTX 150 nmoL/kg, ⑦ 2PTX group, injected with PTX 300 nmol/kg, and ⑧2(PTX-OCT) injected with PTX-OCT 300 nmol/kg, The tumor volume and body weight (BW) were observed regularly. The tumor volume doubling time was calculated. White blood cells were counted by collecting peripheral blood sample. By the end of experiment the mice were killed with the tumors taken out. The expression of mRNA of subtypesl-5 of human somatostatin receptor (SSTR1-SSTR5 ) were investigated using RT-PCR Histological apoptosis was detected by DNA ladder. Immunohistoehemistry was used to examine the SSTR2 and SSTR5 expression and tumor microvessel density (MVD). Results The tumor volumes of 2PTX-OCT and 2 (2PTX-OCT) groups were significantly smaller than those of other groups (all P< 0.01 ). The tumor doubling times of the 2tPTX-OCT and 2(2PTX-OCT) groups were significantly longer than those of the other groups too (al.1P<0.01). The MVD levels of the 2tTX-OCT and 2(2PTX-OCT) groups were significant lower than those of the other groups (all P<0.01 ). The toxicity of the PTX group was more obvious. The WBC count levels of the PTX and 2PTX groups were significantly lower than those of the other groups ( all P< 0.05 ). mRNA expression could be found for SSTR1, 2, 4, and 5 in the A549 xenngrafts. Immunohistochemistry showed that SSTR2 was maiuly found in the tumor cell membrane, and $STR5 was found in the tumor cell membrane and cytoplasm. More histological apoptosis bands were shown by DNA ladder method in the 2PTX-OCT and 2 (PTX-OCT) groups. Conclusion The targeting conjugate 2PTX-OCT inhibits the proliferation of tumor ceils, reduces microvessel, and decreases the toxicity in comparison with the cytotoxic radical PTX.