摘要
目的 探讨长链非编码RNA(lncRNA)C3orf77-2对头颈鳞癌细胞增殖和迁移的影响.方法 收集2021年9月至2023年9月郑州大学第一附属医院收治并接受手术治疗的30例下咽鳞癌患者的临床样本作为研究对象.选取下咽鳞癌组织和癌旁组织标本,运用lncRNA芯片技术检测下咽鳞癌组织和配对的癌旁组织标本中lncRNA的表达,分析差异表达的非编码RNA;实时定量反转录聚合酶链反应(RT-qPCR)验证基因芯片差异基因;收集临床数据,分析lncRNA C3orf77-2表达与各种临床病理的相关性;下咽鳞癌FaDu细胞随机分为对照组、siNC组、lncRNA C3orf77-2 si-lnc1组、lncRNA C3orf77-2 silnc2组;沉默lncRNA C3orf77-2后,采用细胞计数试剂盒(CCK-8)和克隆形成实验检测细胞增殖能力,细胞划痕实验检测细胞迁移能力;组间比较采用最小显著差异法LSD-t检验进行统计分析.结果 下咽鳞癌组织中lncRNA C3orf77-2表达量(0.78±0.14)显著高于癌旁组织(0.13±0.10),差异有统计学意义(t=14.96,P＜0.01).下咽鳞癌组织中lncRNA C3orf77-2的表达与患者肿瘤分化程度(F=4.3,P＜0.05)、TNM分期(F=13.4,P＜0.05)、淋巴结转移((=4.07,P＜0.05)明显相关;CCK-8结果显示,各组细胞在24、48、72 h的吸光度值分别为空白对照组(0.360±0.018、0.546±0.047、0.882±0.028),siNC 组(0.401±0.018、0.561±0.037、0.825±0.056),ln-cRNA C3orf77-2 si-lnc1(0.230±0.031、0.416±0.009、0.555±0.048)和lncRNA C3orf77-2 si-lnc2(0.244±0.008、0.367±0.041、0.531±0.022),差异有统计学意义(F24h=15.6、F48h=360.7、F72h=105.7,P＜0.01).克隆形成实验结果显示,siNC 组(89.00±0.58),lncRNA C3orf77-2 si-lnc1(31.67±1.76),lncRNA C3orf77-2 si-lnc2(22.67±1.76),空白对照组(94.00±2.52),差异有统计学意义(F=433.6,P＜0.01).细胞划痕实验显示:si-NC 组(36.67±4.33),lncRNA C3orf77-2 si-lnc1(10.83±1.17),lncRNA C3orf77-2 si-lnc2(12.00±1.67),差异有统计学意义(F=78.8,P＜0.01).结论 lncRNA C3orf77-2在下咽鳞癌组织中表达显著上调,且与患者的临床病理特征相关.沉默lncRNA C3orf77-2可显著抑制FaDu细胞的增殖、迁移.
Abstract
Objective To investigate the effect of long chain non-coding RNA(lncRNA)C3orf77-2 on the proliferation and migration of head and neck squamous carcinoma cells.Methods Clinical samples from 30 patients with squamous carcinoma of the hypopharynx admitted to the First Affiliated Hospital of Zhengzhou University and underwent surgical treatment from September 2021 to September 2023 were collected as study subjects.Hypopharyngeal squamous carcinoma tissues and paracancerous tissue speci-mens were selected,and lncRNA microarray technology was used to detect the expression of long-chain noncoding RNAs in hypopharyngeal squamous carcinoma tissues and paired paracancerous tissue speci-mens,and to analyze the differentially expressed noncoding RNAs;RT-qPCR was performed to validate the differential genes of gene microarray;and the clinical data were collected and analyzed for the correlation between the expression of lncRNA C3orf77-2 and various clinicopathologies;Hypopharyngeal squamous carcinoma FaDu cells were randomly divided into control group,siNC group,lncRNA C3orf77-2 si-lncl group,lncRNA C3orf77-2 silnc2 group;after silencing of lncRNA C3orf77-2,cell proliferation ability was detected by cell counting kit-8(CCK-8)and clone formation assay,and cell migration ability was detected by cell scratch assay;the comparison between groups was made by the least significant difference method LSD;the comparison between groups was made by the least significant difference method LSD.Compari-sons between groups were statistically analyzed using the least significant difference method LSD-t test.Results The expression of lncRNA C3orf77-2 in hypopharyngeal squamous carcinoma tissues(0.78±0.14)was significantly higher than that in paracancerous tissues(0.13±0.10),and the difference was statistically significant(t=14.96,P＜0.01).The expression of lncRNA C3orf77-2 in hypopharyngeal squamous carcinoma tissues was significantly correlated with the degree of tumor differentiation(F=4.3,P＜0.05),TNM stage(F=13.4,P＜0.05),and lymph node metastasis(t=4.07,P＜0.05)in the pa-tients;the results of CCK-8 showed that the absorbance values of the cells of each group at 24,48,72 h were blank control group(0.360±0.018,0.546±0.047,0.882±0.028),siNC group(0.401±0.018,0.561±0.037,0.825±0.056),and lncRNA C3orf77-2 si-lnc1(0.23±0.031,0.416±0.009,0.555±0.048)and lncRNA C3orf77-2 si-lnc2(0.244±0.008,0.367±0.041,0.531±0.022),and the difference was statistically significant(F24 h=15.6,F48 h=360.7,F72 h=105.7,P＜0.01).The results of clone formation experiments showed that the siNC group(89.00±0.58),lncRNA C3orf77-2 si-lnc1(31.67±1.76),lncRNA C3orf77-2 si-lnc2(22.67±1.76),and the blank control group(94.00±2.52),with a statistically significant difference(F=433.6,P＜0.01).Cell scratch experiment showed that:si-NC group(36.67±4.33),lncRNA C3orf77-2 si-lnc1(10.83±1.17),lncRNA C3orf77-2 si-lnc2(12.00±1.67),and the difference was statistically significant(F=78.8,P＜0.01).Conclusion The expression of lncRNA C3orf77-2 was significantly upregulated in hypopharyngeal squamous carcinoma tissues and cor-related with the clinicopathological features of patients.Silencing of lncRNA C3orf77-2 significantly inhibi-ted the proliferation and migration of FaDu cells.
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