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目的:探讨长链非编码RNA（long non-coding RNA，LncRNA）AC000061.1调节n CFTR基因表达在非梗阻性无精子症（nonobstructive azoospermia，NOA）发病机制中的作用。n 方法:基因芯片检测梗阻性无精子症（obstructive azoospermia，OA）组（50例）、NOA组（50例）及对照组（50例）患者的睾丸组织中差异表达的LncRNA 并对其对应的靶向mRNA进行生物信息学分析，用qRT-PCR和Western blotting法检测三组患者的睾丸组织凋亡基因n Bcl-2表达差异。qRT-PCR验证三组患者睾丸组织、血清、精浆中LncRNA AC000061.1和n CFTR mRNA表达水平。酶联免疫吸附法（enzyme-linked immunosorbent assay，ELISA）检测血清和精浆中CFTR蛋白浓度。构建LncRNA AC000061.1过表达（H-LncRNA组）、沉默（Si-LncRNA组）及空载对照组载体转染至睾丸癌细胞系（NTERA-2），qRT-PCR验证LncRNA AC000061.1及n CFTR mRNA的表达，Western blotting检测CFTR蛋白水平，CCK-8检测细胞增殖能力，流式细胞术及TUNEL凋亡试剂盒检测细胞凋亡率。n 结果:芯片检测和qRT-PCR显示，与对照组相比，NOA组睾丸组织LncRNA AC000061.1和n CFTR表达降低（n P=0.033，n P=0.042），OA组LncRNA AC000061.1表达无差异，n CFTR低表达（n P=0.039）；OA组和NOA组凋亡基因n Bcl-2表达水平依次增高（n P=0.031，n P=0.008）。三组血清中LncRNA AC000061.1 mRNA和n CFTR mRNA及蛋白表达水平差异均无统计学意义（n P均>0.05）。在精浆中，与对照组相比，NOA组和OA组LncRNA AC000061.1和n CFTR mRNA及蛋白含量依次降低（n P=0.002，n P=0.038和n P=0.006，n P=0.026），且LncRNA AC000061.1与n CFTR mRNA呈正相关（n r=0.169，n P=0.039）。质粒转染NTERA-2细胞后，沉默Si-LncRNA组LncRNA AC000061.1 mRNA及n CFTR mRNA和蛋白表达均降低（n P=0.005，n P=0.003），过表达H-LncRNA组LncRNA AC000061.1 mRNA及n CFTR mRNA和蛋白表达均显著增高（n P=0.002，n P=0.009）。沉默Si-LncRNA组细胞增殖能力显著下降（n P=0.003），细胞凋亡率明显增高（n P=0.001）；过表达H-LncRNA组细胞增殖能力增加（n P=0.017），细胞凋亡率降低（n P=0.017）。n 结论:NOA睾丸组织中LncRNA AC00006.1通过调控n CFTR基因表达引发细胞增殖与凋亡的异常，可能参与NOA的发病机制。n “,”Objective:To explore the role of long non-coding RNA (LncRNA) AC000061.1 involved in the pathogenesis of nonobstructive azoospermia (NOA) by regulating the n CFTR gene expression.n Methods:Bioinformatics analysis was conducted with the use of gene microarray to screen the differentially expressed LncRNAs and corresponding mRNAs in the testicular tissue of patients in three groups including obstructive azoospermia (OA) group (n n=50), NOA group (n n=50), and control group (n n=50). The expression of apoptosis-related gene n Bcl-2 in the testicular tissue of patients in these three groups was detected by qRT-PCR and Western blotting. Additionally, qRT-PCR was performed to determine the expression of LncRNA AC000061.1 and n CFTR mRNA in the testicular tissue, serum, and seminal plasma of patients in the three groups, and enzyme-linked immunosorbent assay (ELISA) was conducted to detect the CFTR protein level in the serum and seminal plasma. LncRNA AC000061.1 overexpression (H-LncRNA group) and silencing vectors (Si-LncRNA group) and empty vectors were constructed and transfected into the testicular cancer cell line NTERA-2. Subsequently, the expression of LncRNA AC000061.1 and n CFTR mRNA was determined by qRT-PCR, and CFTR protein expression was measured by Western blotting assay. Cell proliferation was assessed using CCK-8 and cell apoptosis was evaluated by flow cytometry and TUNEL.n Results:Microarray analysis and qRT-PCR showed that the expression of LncRNA AC006100.1 and CFTR decreased in NOA group compared with control group (n P=0.033 and n P=0.042). But there was no difference in the expression of LncRNA AC000061.1 between OA group and control group, while CFTR in OA group was lowly expressed compared with that in control group (n P=0.039). Bcl-2 expression was sequentially upregulated in OA and NOA groups relative to normal control group (n P=0.031 and n P=0.008). No significant difference was noted in the serum levels of LncRNA AC000061.1 and n CFTR mRNA and protein among the three groups (n P>0.05). The results also showed sequentially decreased levels of LncRNA AC000061.1 andn CFTR mRNA and protein in seminal plasma in NOA and OA groups when compared with control group (n P=0.002, n P=0.038 and n P=0.006, n P=0.026) and a positive correlation of LncRNA AC000061.1 expression with n CFTR mRNA expression (n r=0.169, n P=0.039). LncRNA AC000061.1 and n CFTR mRNA and protein expression decreased in Si-LncRNA group (n P=0.005, n P=0.003), but significantly increased in H-LncRNA group (n P=0.002, n P=0.009). Furthermore, cell proliferation was significantly repressed while apoptosis rate was elevated in Si-LncRNA group (n P=0.003 and n P=0.001). On the contrary, enhanced cell proliferation and inhibited apoptosis were observed in H-LncRNA group (n P=0.017 and n P=0.017).n Conclusion:LncRNA AC00006.1 in the testicular tissue of NOA patients induced abnormal cell proliferation and apoptosis n via mediating n CFTR gene expression, thereby participating in the pathogenesis of NOA.n