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目的:通过对不同质量浓度的磁性抗体体外转染结肠癌干细胞后的系列研究,探索最佳转染浓度。方法:将磁性抗体含铁浓度分别设定为5、10、25、50、75、100 mg/L,分别转染结肠癌干细胞,采用普鲁士蓝染色观察细胞内铁颗粒,原子吸收光谱仪法测定细胞铁含量并检测细胞活性,MR成像检测细胞信号强度变化。结果:各组细胞内铁含量组间比较采用单因素方差分析,组间差异具有统计学意义(F=71.709,P=0.000),在α=0.05的检验水准下,可以认为各组间均数不完全相同,进一步采用单因素方差分析LSD法(双侧检验)进行组间均数多重比较,结果显示,50 mg/L组与5 mg/L组、10 mg/L组、25 mg/L组相比,差异均有统计学意义(P=0.000,P=0.000,P=0.000),但与75 mg/L组、100 mg/L组比较,差异均无统计学意义(P=0.607,P=0.419)。细胞活性检测组间比较采用单因素方差分析,组间差异具有统计学意义(F=69.306,P=0.000),在α=0.05的检验水准下,可以认为各组间均数不完全相同,进一步采用单因素方差分析LSD法(双侧检验)进行组间均数多重比较,结果显示,50 mg/L组与75mg/L比较,细胞活性差异无统计学意义(P=0.134),与5 mg/L组、10 mg/L组、25 mg/L组、100 mg/L组比较,差异均有统计学意义(P=0.000,P=0.000,P=0.000,P=0.008)。T2WI信号强度采用单因素方差分析进行组间比较,组间差异具有统计学意义(F=3 180.748,P=0.000),在α=0.05的检验水准下,可以认为各组间均数不完全相同,进一步采用单因素方差分析LSD法(双侧检验)进行组间均数多重比较,结果显示,50 mg/L组与75 mg/L、100 mg/L各组信号强度之间差异无统计学意义(P=0.945,P=0.960),与空白对照组、细胞对照组、5 mg/L、10 mg/L、25 mg/L各组信号强度相比,差异均有统计学意义(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000)。由此可见,磁性抗体含铁浓度在一定范围内(5~25 mg/L)增加,结肠癌干细胞内铁含量逐渐增加,当磁性抗体含铁浓度为50 mg/L时达到饱和,且细胞活性下降,MR成像T2×WI序列图像显示信号强度明显降低。结论:当磁性抗体含铁浓度为50 mg/L时,可有效转染结肠癌干细胞,能被MR成像可视化。
OBJECTIVE: To explore the optimal transfection concentration by in vitro transfection of colon cancer stem cells with different mass concentrations of magnetic antibodies. Methods: Ferritin concentration was set at 5, 10, 25, 50, 75 and 100 mg / L, respectively, and transfected into colon cancer stem cells. Prussian blue staining was used to observe intracellular iron particles, and cell viability was measured by atomic absorption spectrometry Iron content was measured and cell viability was measured. MR imaging was used to detect cell signal intensity. Results: The intra-articular iron content in each group was compared by one-way ANOVA, the difference between the two groups was statistically significant (F = 71.709, P = 0.000). Under the test level of α = 0.05, The results of multiple comparison among groups showed that 50 mg / L group, 5 mg / L group, 10 mg / L group, 25 mg / L (P = 0.000, P = 0.000, P = 0.000). However, there was no significant difference between the two groups (P = 0.607, P = 0.419). The single-factor analysis of variance (ANOVA) was used to compare the cell viability between the two groups. The difference between the two groups was statistically significant (F = 69.306, P = 0.000). Under the test condition of α = 0.05, the mean between groups was not completely the same The single-factor analysis of variance LSD (two-sided test) for multiple comparison of mean between groups, the results showed that 50 mg / L group compared with 75mg / L, cell activity was no significant difference (P = 0.134), and 5 mg / L group, 10 mg / L group, 25 mg / L group and 100 mg / L group. The difference was statistically significant (P = 0.000; P = 0.000; P = 0.000, P = 0.008). The signal intensity of T2WI was analyzed by one-way ANOVA. The differences between groups were statistically significant (F = 3 180.748, P = 0.000). Under the test level of α = 0.05, the mean between groups was not completely the same , Further using one-way analysis of variance LSD (two-sided test) multiple comparison between the mean number of groups, the results showed that 50 mg / L group and 75 mg / L, 100 mg / L signal intensity of each group no statistical difference (P = 0.945, P = 0.960). Compared with the blank control group, cell control group, 5 mg / L, 10 mg / L and 25 mg / L groups, the difference was statistically significant (P = 0.000, P = 0.000, P = 0.000, P = 0.000, P = 0.000). Thus, the iron content of magnetic antibody increased within a certain range (5 ~ 25 mg / L), the iron content of colon cancer stem cells increased gradually, when the iron content of magnetic antibody reached 50 mg / L saturation, and cell viability Decreased, MR imaging T2 × WI sequence images showed a significant reduction in signal intensity. CONCLUSION: Colon cancer stem cells can be efficiently transfected when magnetic iron concentration is 50 mg / L and can be visualized by MR imaging.