摘要
目的通过体内外实验研究人源化抗表皮生长因子受体(EGFR)单抗尼妥珠(h-R3)对耐多西紫杉醇(DTX)人肺腺癌细胞株(SPC-A1/DTX)化疗敏感性的调变作用。方法免疫组化法及流式细胞仪测定人肺腺癌细胞株SPC-A1和SPC-A1/DTX肺腺癌细胞株表面EGFR的表达强度,突变富集-液相芯片法检测其EGFR、K-Ras和PI3KCA基因的突变情况,流式细胞仪检测h-R3对细胞周期的影响以及h-R3联合DTX对细胞凋亡的影响,MTT法检测h-R3与DTX的联合指数(CI),裸鼠SPC-A1/DTX移植瘤模型观察联合组及单药组对裸鼠抑制瘤模型肿瘤的增殖情况并计算瘤重抑制率(TWI)。结果 SPC-A1细胞株EGFR表达强度为(+,21.53%),SPC-A1/DTX细胞株为(+++,92.47%);SPC-A1/DTX细胞株的PI3KCA基因外显子20突变,SPC-A1细胞株无突变。h-R3作用24h后,SPC-A1/DTX细胞G1期阻滞较SPC-A1细胞更显著(P=0.0002);h-R3及DTX联合用药后SPC-A1/DTX细胞株的凋亡率为(24.7±0.5)%,明显高于h-R3单药组的(14.5±0.1)%,而单用DTX后的凋亡率无显著升高,SPC-A1细胞株单药组及联合组凋亡率均有升高(P<0.05);100μg/ml及200μg/ml h-R3与DTX联合对SPC-A1或SPC-A1/DTX细胞株的增殖抑制具有协同作用;联合组对SPC-A1/DTX肺腺癌荷瘤裸鼠的TWI为71.7%,高于DTX组的52.6%和h-R3组的36.9%。结论 h-R3能明显增加耐DTX的人肺腺癌细胞株化疗的敏感性,其机制可能为h-R3具有G1期阻滞和逆转耐药细胞凋亡抵抗的作用,并且其疗效与耐药细胞较亲本细胞的EGFR表达增强有关,而耐药细胞的PI3KCA基因突变对疗效的影响不显著。
Objective To investigate the effect of nimotuzumab（h-R3） alone or combined with chemotherapy on human adenocarcinoma docetaxel-resistant cell line（SPC-A1/DTX）.Methods The epidermal growth factor receptor（EGFR） status of SPC-A1 and SPC-A1/DTX cell lines were detected by immunohistochemisty and flow cytometry.A mutant-enriched liquid-chip was used in detection of EGFR,K-Ras and PI3KCA mutations.Cell cycle distribution and apoptosis were detected to evaluate effects of h-R3.The growth inhibition was detected by MTT assay and the combination index（CI） was accounted.In vivo,SPC-A1/DTX cells were injected subcutaneously into nude mice,following up tumor size and weight.Results The status of EGFR in SPC-A1was（＋,21.53%）,while in SPC-A1/DTX was（＋＋＋,92.47%）.SPC-A1 was without mutation,and SPC-A1/DTX showed PI3KCA/E20 mutation.An obvious shift into the G1 phase of the cell cycle after 24 hours of 100μg/ml h-R3 treatment（SPC-A1： P=0.018;SPC-A1/DTX： P=0.0002）.In SPC-A1 cell line,apoptosis enhancement effect was revealed in all groups（P0.05）,but in SPC-A1/DTX cell apoptosis enhancement effect was revealed obviously in combined group and single h-R3 group（P0.01）.DTX combined with100μg/ml or 200μg/ml h-R3,cooperative growth inhibitory effect was observed in SPC-A1（CI=0.71） and SPC-A1/DTX（CI=0.40）.h-R3 combined with DTX in nude mice bearing SPC-A1/DTX tumor had stronger suppressive effect on the tumor growth in vivo.Conclusion h-R3 can enhance the chemotherapy effects on the SPC-A1/DTX cell line which may be associated with apoptosis induction and cell cycle arrest at G1 phase.The cooperative growth inhibitory effects may depend on the expression of EGFR other than the mutations of PI3KCA.
出处
《临床肿瘤学杂志》
CAS
2011年第4期298-303,共6页
Chinese Clinical Oncology
基金
南京军区南京总医院青年基金资助项目(Q2008027)
