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目的研究microRNA-214对不同种类淋巴瘤细胞耐药性的影响。方法选择几种不同的淋巴瘤细胞,分别命名为Jurkat细胞实验组、OCI-Ly3细胞实验组及Raji细胞实验组,将健康人外周血淋巴细胞(Scr)作为对照组。细胞分别被敲除miR-214并给予不同的药物刺激。用荧光定量PCR法检测不同淋巴瘤细胞内miR-214的表达量;以CCK-8法检测敲除miR-214后的淋巴瘤细胞存活率;以MTT比色法检测敲除miR-214后不同淋巴瘤细胞对不同药物的敏感性;以蛋白质免疫印迹法检测BCL11B蛋白表达水平;用荧光素酶报告技术检测BCL11B是否为miR-214的靶基因。结果 Jurkat、OCI-Ly3及Raji细胞在实验组的miR-214的表达量分别为27.21±1.37,17.13±2.11,19.31±2.46,均明显高于对照组的1.17±0.28,差异有统计学意义(均P<0.05)。敲除miR-214后,与对照组的Scr细胞存活率(86±12)%相比,Jurkat、OCI-Ly3及Raji细胞在实验组的细胞存活率分别为(42±13)%,(65±11)%,(58±7)%,差异有统计学意义(均P<0.05)。抑制miR-214表达后,对照组Scr细胞对阿霉素、环磷酰胺、柔红霉素、丝裂霉素C和长春新碱的耐药敏感性分别为0.46±0.11,1.96±0.13,1.15±0.22,3.90±0.52,0.52±0.15,Jurkat细胞对5种药物的耐药敏感性分别为0.32±0.07,0.76±0.16,0.73±0.13,2.36±0.16,0.37±0.06,OCI-Ly3细胞对5种药物的耐药敏感性分别为0.28±0.06,0.91±0.12,0.55±0.09,2.01±0.36,0.23±0.07,Raji细胞对5种药物的耐药敏感性分别为0.36±0.04,0.82±0.08,0.64±0.07,3.31±0.21,0.35±0.05,与对照组比较,差异有统计学意义(P<0.05)。BCL11B是miR-214的靶基因。结论抑制miR-214表达能够降低不同种类淋巴瘤细胞的存活率,并降低白血病淋巴细胞的多药耐药性,这种作用可能与miR-214抑制其靶基因BCL11B的表达相关。
Objective To study the effect of microRNA-214 on the drug resistance of different kinds of lymphoma cells. Methods Several different lymphoma cells were selected and named as Jurkat cells experimental group, OCI-Ly3 cells experimental group and Raji cells experimental group, and healthy human peripheral blood lymphocytes (Scr) as control group. Cells were knocked out of miR-214 and given different drug stimuli. The expression of miR-214 in different lymphoma cells was detected by real-time PCR. The survival rate of lymphoma cells after knocking down miR-214 was detected by CCK-8 assay. The expression of miR-214 was detected by MTT assay Lymphoma cells to different drugs sensitivity; Western blot detection of BCL11B protein expression; using luciferase reporter assay to detect whether BCL11B miR-214 target gene. Results The expression of miR-214 in Jurkat, OCI-Ly3 and Raji cells in experimental group were 27.21 ± 1.37, 17.13 ± 2.11 and 19.31 ± 2.46, respectively, which were significantly higher than those in control group (1.17 ± 0.28, respectively) All P <0.05). After knocking down miR-214, the survival rates of Jurkat, OCI-Ly3 and Raji cells in the experimental group were (42 ± 13)% and (65 ± 11%, (58 ± 7)%, respectively, with significant difference (all P <0.05). The sensitivity of Scr cells to doxorubicin, cyclophosphamide, daunorubicin, mitomycin C and vincristine was 0.46 ± 0.11, 1.96 ± 0.13 and 1.15 respectively after inhibiting miR-214 expression ± 0.22, 3.90 ± 0.52 and 0.52 ± 0.15 respectively. The sensitivity of Jurkat cells to five drugs were 0.32 ± 0.07,0.76 ± 0.16,0.73 ± 0.13,2.36 ± 0.16,0.37 ± 0.06, respectively. The sensitivity of OCI-Ly3 cells to 5 The susceptibilities of these drugs to drug resistance were 0.28 ± 0.06,0.91 ± 0.12,0.55 ± 0.09,2.01 ± 0.36,0.23 ± 0.07 respectively. The sensitivity of Raji cells to the five drugs were 0.36 ± 0.04,0.82 ± 0.08, 0.64 ± 0.07,3.31 ± 0.21,0.35 ± 0.05, the difference was statistically significant (P <0.05) compared with the control group. BCL11B is the target gene for miR-214. Conclusion Inhibition of miR-214 expression can reduce the survival rate of different types of lymphoma cells and reduce the multidrug resistance of leukemia lymphocytes, which may be related to the inhibition of miR-214 expression by its target gene BCL11B.