长链非编码 SNHG15靶向 miR-141对甲状腺癌细胞侵袭及凋亡的影响研究

目的 探讨长链非编码(lncRNA)核仁小RNA宿主基因15(SNHG15)靶向调节微小RNA-141(miR-141)对甲状腺癌细胞侵袭、凋亡影响及机制.方法 本研究起止时间为2018年10月至2019年5月,参照Lipofectamine 2000说明将SNHG15小干扰RNA(siRNA)、siRNA阴性对照(siRNA control)、miR-141抑制剂(miR-141 inhibitor)或抑制剂阴性对照(inhibitor control)转染至甲状腺癌FRO细胞,细胞随机分组空白对照(si-control)组、转染阴性对照(si-NC)组、转染SNHG15 siRNA(si-SNHG15)组、转染SNHG15 siRNA和inhibitor control(si-SNHG15+anti-control)组和转染SNHG15 siRNA和miR-141 inhibitor(si-SNHG15+anti-miR-141)组,,转染48 h,qRT-PCR检测SNHG15和miR-141 mRNA表达;Transwell小室及流式细胞术分别检测细胞侵袭能力及凋亡率;双荧光素酶报告系统检测SNHG15和miR-141的靶向关系.Western blotting检测上皮钙黏素(E-cadherin)、B细胞淋巴瘤/白血病-2(Bcl-2)和Bcl-2相关X蛋白(Bax)蛋白表达.结果 与si-control组比较,si-SNHG15组中SNHG15 mRNA表达[(1.000)比(0.263±0.032)]明显降低,细胞侵袭能力[(168.6±7.1)个比(78.2±3.3)个]明显降低,凋亡率[(4.31±0.42)％比(33.11±1.69)％]明显升高,E-cadherin[(0.105±0.011)比(0.602±0.058)]和Bax表达[(0.049±0.008)比(0.263±0.028)]明显升高,Bcl-2表达[(0.587±0.063)比(0.131±0.015)]明显降低(P<0.05).SNHG15与miR-141存在靶向关系.与si-SNHG15+anti-control组比较,si-SNHG15+anti-miR-141组细胞侵袭能力[(79.9±4.2)个比(130.3±5.2)个]明显升高,凋亡率[(34.08±1.63)％比(15.66±0.87)％]降低,E-cadherin[(0.641±0.062)比(0.309±0.032)]和Bax表达[(0.282±0.030)比(0.144±0.015)]明显降低,Bcl-2表达[(0.138±0.017)比(0.478±0.052)]明显升高(P<0.05).结论 lncRNA SNHG15可靶向调节miR-141影响甲状腺癌细胞侵袭和凋亡,机制与调节E-cadherin、Bcl-2和Bax表达有关.

Effects of long-chain non-coding SNHG15 targeting miR-141 on invasion and apoptosis of thyroid cancer cells

Objective To explore the effect of long-chain non-coding RNA (lncRNA) small nucleolar RNA host gene 15 (SNHG15) targeting miR-141 on invasion and apoptosis of thyroid cancer and its mechanism.Methods This research started from October 2018 and ended in May 2019. According to Lipofectamine 2000, SNHG15 small interfering RNA (siRNA), siRNA negative control (siRNAcontrol), miR-141 inhibitor (miR-141 inhibitor) or Inhibitor negative control (inhibitor control) was transfected into thyroid cancer FROcells. The cells were randomly assigned into blank control (si-control) group, transfection negative control (si-NC) group, transfection of SNHG15 siRNA (si-SNHG15) group, transfection of SNHG15 siRNA and inhibitor control (si-SNHG15+anti-control) group and transfection of SNHG15 siRNA and miR-141 inhibitor (si-SNHG15+anti-miR-141) group. Cells were transfected for 48 hours, and the expressions of SNHG15 and miR-141 were detected by qRT-PCR. The invasive ability and apoptotic rate of cells were detected by Transwell chamber and flow cytometry. The targeting relationship between SNHG15 and miR-141 was detected by double luciferase reporting system. Western blotting was used to detect the expressions of E-cadherin, B-cell lymphoma/leukemia-2 (Bcl-2) and Bcl-2 related X protein (Bax) proteins.Results In the si-SNHG15 group, by comparison with the si-control group, the expressions of SNHG15 mRNA [(1.000) vs (0.263±0.032)] and Bcl-2 [(0.587±0.063) vs (0.131±0.015)] and the cell invasion ability [(168.6±7.1) vs (78.2±3.3)] were significantly lower, , while the apoptotic rate [(4.31±0.42) % vs (33.11±1.69)%], and the expressions of E-cadherin [(0.105±0.011) vs (0.602±0.058)] and Bax [(0.049±0.008) vs ( 0.263±0.028)] were significantly higher (P<0.05). SNHG15 had a targeting relationship with miR-141. In the si-SNHG15+anti-miR-141 group, by comparison with the si-SNHG15+anti-control group, the cell invasion ability [(79.9±4.2) vs (130.3±5.2)] and Bcl-2 expression [(0.138±0.017) vs (0.478±0.052)] were significantly higher, while the apoptosis rate [(34.08± 1.63)% vs (15.66±0.87)%], and the expressions of E-cadherin [(0.641±0.062) vs (0.309±0.032)] and Bax [(0.282±0.030) vs (0.144±0.015)] were significantly lower (P<0.05).Conclusion lncRNA SNHG15 can target miR-141 to affect the invasion and apoptosis of thyroid cancer cells, and its mechanism is related to regulating the expressions of E-cadherin, Bcl-2 and Bax.