摘要
目的耐药是影响宫颈癌治疗效果和预后的重要因素,尼妥珠单抗为我国首个功能性肿瘤治疗单克隆抗体,尼妥珠单抗、调强放疗、顺铂能够从不同方面达到杀伤肿瘤细胞的目的,但是关于三者联合治疗耐药性宫颈癌的临床和实验研究鲜有报道。本研究通过动物实验,探讨尼妥珠单抗联合调强放疗及顺铂治疗耐药性宫颈癌的治疗效果和联合治疗机制,为宫颈癌联合治疗提供帮助。方法建立顺铂耐药性HeLa细胞系,利用该细胞系建立宫颈癌裸鼠模型,并随机分为4组,每组15只。对照(Control)组荷瘤裸鼠给予磷酸盐缓冲液治疗,放化疗(RTCT)组荷瘤裸鼠给予调强放疗,顺铂治疗、尼妥珠单抗(H-R3)组荷瘤裸鼠给予尼妥珠单抗治疗,RTCT+H-R3组荷瘤裸鼠依次给予尼妥珠单抗、顺铂和调强放疗治疗。定期测定各组荷瘤裸鼠肿瘤长、短直径和平均存活时间;治疗22d,断颈处死后收集肿瘤组织进行苏木精-HE染色同时蛋白质印迹法检测肿瘤组织Akt、p-Akt、MEK及p-MEK蛋白表达情况。结果各组的肿瘤体积比均随治疗时间变化,且变化趋势有较明显不同,组间维度(F组间=24.103,P<0.001)、时间度(F时间=15.510,P<0.001)和交互作用维度(F交互=3.592,P=0.020),差异有统计学意义。进一步采取两两比较发现,RTCT+H-R3组荷瘤裸鼠经尼妥珠单抗和放化疗联合治疗22d后,肿瘤体积比为(3.38±0.71),显著低于Control组(17.21±1.35)、RTCT组(14.56±1.14)和H-R3组(10.79±1.08),差异有统计学意义,P<0.05。同时RTCT+H-R3组荷瘤裸鼠平均存活时间为66d,明显高于Control组的28d、RTCT组的37d和H-R3组的49d,依次达到2.36、1.78和1.35倍。HE染色结果表明,RTCT+H-R3组荷瘤裸鼠细胞核出现严重固缩,同时肿瘤组织出现空泡现象,病理学改变最明显;蛋白质印迹法表明,RTCT+H-R3组Akt(0.34±0.05)、p-Akt(0.15±0.02)、MEK(0.27±0.03)和p-MEK(0.23±0.02)表达量最低,其次为H-R3组,Control组和RTCT组Akt、p-Akt、MEK、p-MEK蛋白表达量相近。结论尼妥珠单抗联合调强放疗及顺铂治疗减缓了顺铂耐药性宫颈癌荷瘤裸鼠肿瘤生长,提高了荷瘤裸鼠存活时间,其机制可能与抑制Akt及MEK有关。
OBJECTIVE Drug resistance is an important factor of influence cervical cancer treatment effect and prognosis, nitozumab is the first functional oncology for monoelonal antibodies. Nitozumab, radiotherapy and cisplatin can achieve the purpose of killing tumor ceils from different aspects, but clinical and experimental research about the com- bined treatment in cervical cancer resistance is rarely reported. This study was conducted through animal experimentsm, to investigate the therapeutic effect and the combined treatment mechanism of the combination of nimotuzumab and inten- sity modulated radiation therapy with cispiatin in the treatment of cervical cancer with drug resistance and to provide as- sistance for the treatment of cervical cancer. METHODS Cisplatin resistant HeLa cell line was established, then it wasused to establish the nude mice model of cervical cancer. The nude mice with cervical cancer were randomly divided into 4 groups, with each group containing 15 mice. The nude mice of control group were treated with PBS; The nude mice of RTCT group were treated with intensity modulated radiation therapy and cisplatin~ The nude mice of H-R3 group were treated with nimotuzumab The nude mice of RTCT H-R3 group were treated with nimotuzumab, eisplatin and intensity modulated radiation therapy. The length, diameter and average survival time of tumor bearing nude mice were measured. After the treatment of 22d, tumor tissues were collected after sacrificed the mouse by the neck and stained with hematox- ylin eosin staining （HE）, and Akt, p-Akt, MEK and p-MEK were detected using Western blot. RESULTS The tumor volume ratio of each group varied with the time of treatment, and the variation trend was significantly different, and there were statistically significant in group （Fi.torblock =24. 103, P〈0. 001）, time（Fmo = 15. 510 ,P〈0. 001_and and interaction （F ~=3. 592, P=0. 020）. after treatment 22d, tumor volume ratio of RTCT±H-R3 group （3.38±0.71） was signifi- cantly lower than that of control group （17.21：kl. 35）, RTCT group （14.56±1.14） and H-R3 group （10.79±1.08） （P〈0.05）. After treatment 22 d, the average survival time of tumor bearing nude mice in group RTCT4-H-R3 was 66 d, which was 2.36, 1.78 and 1.35 times of control group（28 d）, RTCT group （37 d） and H R3 group （49 d）, re- spectively. The results of HE staining showed that the pathological changes of tumor bearing nude mice in RTCT H-R3 group were the most obvious,the nuclei appeared severe shrinkage, while the tumor tissue appeared cavitation. The re- sults of Western blot showed that the expressions of Akt （0.34±0.05）, p-Akt （0.15±0.02）, MEK （0.27±0.03） and p-MEK （0. 23±0. 02） were the lowest in RTCT H-R3 group, followed by H-R3 group, but Akt, p-Akt, MEK, p-MEK protein expressions in control group and RTCT group were similar. CONCLUSIONS The combination of nimotu- zumab and intensity modulated radiation therapy with cisplatin decreased the growth of tumor in nude mice and improved the survival time. Its mechanism may be related to Akt and MEK.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2017年第18期1284-1289,共6页
Chinese Journal of Cancer Prevention and Treatment
关键词
尼妥珠单抗
调强放疗
顺铂
联合治疗
宫颈癌
Nimotuzumab
intensity modulated radiation therapy
Cisplatinl combination therapy
cervical cancer
