摘要
目的探讨人端粒酶逆转录酶反义寡核苷酸(hTERT ASODN)对乳腺癌MCF-7细胞系干细胞微球体富集及其生物学特性的影响。方法以人端粒酶逆转录酶(hTERT)基因编码区为靶点合成反义寡核苷酸,脂质体介导转染,以正义序列转染作为对照。实验分设转染组(ASODN组),正义序列转染的对照组(SODN组)及空白对照组。采用无血清悬浮培养法观察转染对MCF-7干细胞微球体富集的影响。Western blot检测各组干性标记物CD44及ALDH1的表达。RT-PCR检测各组ABCG2、MDR1 mRNA的表达。采用单因素方差分析对各组数据进行统计学处理。结果 ASODN组干细胞微球体生长速率及球体体积明显差于两对照组。与空白对照及SODN组比较,Western blot检测发现CD44与ALDH1在ASODN组细胞表达降低,差异有统计学意义(P<0.01)。RT-PCR检测发现,在ASODN组细胞ABCG2及MDR1 mRNA的表达量均显著降低,差异有统计学意义(P<0.01)。而空白对照组及SODN组两者比较差异无统计学意义(P>0.05)。结论 hTERT ASODN在抑制乳腺癌干细胞微球体的富集、降低乳腺癌细胞干性表达及其耐药性方面具有重要作用。
Objective To investigate the effects of telomerase reversetranscriptase antisense oligonucleotide（hTERT-ASODN） on stem cell microsphere enrichment and biochemical characteristics of breast cancer MCF-7 cell. Methods Antisense oligonucleotides according to hTERT target gene coding region was synthesized for liposome-mediated transfection. Sense oligodeoxynucleotide transfection was treated as control. Set up ASODN transfection group, SODN transfection group and blank control group respectively. The effect of ASODN transfection on MCF-7 stem cell microsphere enrichment was tested by serum-free medium Suspension culture. The expressions of stem cell markers CD44 and ALDH1 were detected by Western blot, the expressions of ABCG2 and MDR1 mRNA in each group were assessed by RT-PCR respectively. The results were analysed by one-way ANOVE, P〈0.05 was considered statistically significant. Results Stem cell microsphere growth rate and sphere volume in ASODN group were significantly worse than which in both control groups. Compared with the blank control and SODN transfection group, there was a lower expression of CD44 and ALDH1 in the ASODN transfection group by Western blot （P〈0.01）. The mRNA＇ expressions of ABCG2 and MDR1 in the ASODN transfection group were significantly lower than which in other groups by RT-PCR （P〈0.01）. There was no significant difference between the blank control and SODN transfection group （P〉0.05）. Conclusion hTERT-ASODN may play an important role in inhibiting the microsphere enrichment of breast cancer stem cells, decreasing the expression of stem cell markers and reducing its drug resistance.
出处
《中华临床医师杂志（电子版）》
CAS
2014年第15期41-44,共4页
Chinese Journal of Clinicians(Electronic Edition)
