LncRNA SNHG1在甲状腺乳头状癌中的表达及其对细胞增殖与凋亡的影响

Expression of lncRNA SNHG1 in papillary thyroid carcinoma and its effect on cell proliferation and apoptosis

目的探讨长链非编码RNA核仁小分子RNA宿主基因(SNHG1)在甲状腺乳头状癌(PTC)组织和细胞中的表达及其对PTC细胞增殖与凋亡的影响。方法采用实时荧光定量PCR(QPCR)检测40例行手术治疗的PTC患者的肿瘤组织和对应癌旁组织中SNHG1的表达水平,同时检测不同PTC细胞系(K1、BCPAP和TPC-1)中SNHG1的表达情况。采用si-SNHG1-2、si-SNHG1-3转染BCPAP和TPC-1细胞,另转染si-NC作为阴性对照组,MTT法检测细胞增殖变化,流式细胞仪检测细胞周期、细胞凋亡变化; Western blotting法检测p53通路相关蛋白的表达变化。结果与癌旁正常组织比较,PTC组织中SNHG1表达增加(2. 72±0. 97 vs. 4. 72±1. 14,P<0. 05); K1、BCPAP和TPC-1细胞中SNHG1表达水平分别为2. 82±0. 25、5. 31±0. 51和3. 92±0. 47,高于人正常甲状腺Nthy-ori3-1细胞(P<0. 05)。与si-NC组比较,si-SNHG1-2和si-SNHG1-3组的BCPAP、TPC-1细胞增殖率分别为(50. 22±4. 65)%和(58. 19±5. 46)%、(47. 18±4. 27)%和(51. 24±6. 49)%,差异均有统计学意义(P<0. 05)。si-NC组BCPAP、TPC-1细胞的G2/M期比例均低于si-SNHG1-2组和si-SNHG1-3组(P<0. 05)。si-NC组BCPAP细胞的凋亡率为(10. 71±0. 78)%,均低于si-SNHG1-2组(24. 90±1. 87)%和si-SNHG1-3组(26. 89±1. 44)%(P<0. 05)。si-NC组TPC-1细胞的凋亡率为(11. 12±1. 05)%,均低于si-SNHG1-2组(27. 21±2. 09)%和si-SNHG1-3组(29. 67±2. 14)%(P<0. 05)。沉默SNHG1表达可降低MDM2蛋白表达并增加p53和p21蛋白表达。结论 SNHG1在PTC组织和细胞中表达均上调;沉默SNHG1表达可能通过激活p53通路来抑制细胞增殖,影响细胞周期并促进凋亡。

Objective To investigate the expression of long-chain non-coding RNA small nucleolar RNA host gene 1( SNHG1)in thyroid papillary carcinoma( PTC) tissues and cells and its effect on the proliferation and apoptosis of PTC cells. Methods Quantitative real-time fluorescence PCR( QPCR) was used to detect the expression of SNHG1 in tumors and adjacent tissues of 40 PTC patients undergoing surgical treatment,and the expression of SNHG1 in different PTC cell lines( K1,BCPAP and TPC-1) was also detected. BCPAP and TPC-1 cells were transfected with si-SNHG1-2 and si-SNHG1-3,and cells transfected with si-NC were used as negative control group. MTT assay was used to detect the changes of cell proliferation. Flow cytometry was used to detect the changes of cell cycle and apoptosis,and Western blotting was used to detect the changes of p53 pathway-related proteins. Results Compared with normal tissues adjacent to cancer,the expression of SNHG1 in PTC tissues increased significantly( 2. 72±0. 97 vs. 4. 72±1. 14,P<0. 05). The expression levels of SNHG1 in K1,BCPAP and TPC-1 cell lines were 2. 82±0. 25,5. 31±0. 51 and 3. 92±0. 47,all higher than that in human normal thyroid Nthy-ori3-1 cells( P < 0. 05). Compared with si-NC group,the proliferative rates of BCPAP cells in si-SNHG1-2 and siSNHG1-3 groups were( 50. 22±4. 65) % and( 58. 19±5. 46) %( P<0. 05),while those of TPC-1 cells in si-SNHG1-2 and si-SNHG1-3groups were( 47. 18±4. 27) % and( 51. 24±6. 49) %( P<0. 05). The G2/M phase ratio of BCPAP and TPC-1 cells in si-NC group was lower than that in si-SNHG1-2 group and si-SNHG1-3 group. The apoptotic rate of BCPAP cells in si-NC group was( 10. 71±0. 78) %,lower than( 24. 90±1. 87) % and( 26. 89±1. 44) % in si-SNHG1-2 group and si-SNHG1-3 group( P<0. 05). The apoptotic rate of TPC-1cells in si-NC group was( 11. 12±1. 05) %,lower than( 27. 21±2. 09) % and( 29. 67±2. 14) % in si-SNHG1-2 group and si-SNHG1-3group( P<0. 05). Silencing the expression of SNHG1 could decrease the expression of MDM2 and increase the expression of p53 and p21.Conclusion SNHG1 expression is up-regulated in PTC tissues and cells. Silencing SNHG1 expression may inhibit cell proliferation,affect cell cycle change and promote apoptosis by activating p53 pathway.