HOTTIP在口腔鳞癌中的表达及意义

Expression and significance of HOTTIP in oral squamous cell carcinoma

目的探讨口腔鳞状细胞癌(OSCC,简称鳞癌)中HOXA远端转录本(HOTTIP)的表达及意义。方法采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)方法检测HOTTIP在正常人口腔黏膜角质形成细胞(HOK)和人OSCC细胞系(HSC-4)中的表达情况;对HSC-4细胞转染HOTTIP-inhibitor,qRT-PCR检测转染后HOTTIP的表达量,MTT实验、细胞划痕实验检测转染后对HSC-4细胞增殖、迁移的影响。结果通过qRT-PCR检测HOTTIP的表达量,HSC-4细胞中的HOTTIP表达量(1.521±0.117)显著高于HOK细胞的(1.060±0.036),差异有统计学意义(P<0.05);对HSC-4细胞转染HOTTIP-inhibitor,qRT-PCR检测HSC-4细胞转染后HOTTIP的表达量,HOTTIP-inhibitor组HOTTIP的表达量(0.336±0.070)均明显低于Control组和Inhibitor-NC组的(1.062±0.092)、(1.023±0.103),差异均有统计学意义(P<0.05);MTT实验证实HSC-4细胞转染HOTTIP-inhibitor后,HOTTIP-inhibitor组相对于Control组和Inhibitor-NC组,细胞增殖能力明显下降,差异有统计学意义(P<0.05),提示敲低HOTTIP的表达能显著降低HSC-4细胞的增殖能力;细胞划痕实验中,HOTTIP-inhibitor组细胞的划痕愈合速度减慢,HOTTIPinhibitor组转染后24 h细胞划痕占比(53.27±1.85)均大于Control组和Inhibitor-NC组的(21.10±0.78)、(25.13±1.69),差异均具有统计学意义(P<0.05),提示HOTTIP的低表达能有效地降低细胞迁移能力。结论与HOK细胞相比,HSC-4细胞中HOTTIP表达明显上调,敲低HOTTIP的表达能显著降低HSC-4细胞的增殖能力、迁移能力,证实其在OSCC中发挥促癌作用。

Objective To discuss the expression and significance of HOXA transcript at the distal tip(HOTTIP)in oral squamous cell carcinoma(OSCC).Methods Real-time fluorescent quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to detect the expression of HOTTIP in normal human oral keratinocytes(HOK)and human OSCC cell line(HSC-4).The expression of HOTTIP after transfection was detected by qRT-PCR,and the effects of transfection on proliferation and migration of HSC-4 cells were detected by MTT assay and cell scratch assay.Results The expression of HOTTIP was detected by qRT-PCR.The expression of HOTTIP in HSC-4 cells(1.521±0.117)was significantly higher than(1.060±0.036)in HOK cells,and the difference was statistically significant(P<0.05).HSC-4 cells were transfected with HOTTIPinhibitor.QRT-PCR was used to detect the expression of HOTTIP in HSC-4 cells.The expression of HOTTIP in the HOTTIP-inhibitor group(0.336±0.070)was significantly lower than(1.062±0.092)and(1.023±0.103)in the Control group and Inhibitor-NC group,their differences were statistically significant(P<0.05).MTT experiments confirmed that after HSC-4 cells were transfected with HOTTIP-inhibitor,the cell proliferation ability of HOTTIP-inhibitor group was significantly lower than that of Control group and Inhibitor-NC group,and the difference was statistically significant(P<0.05),suggesting that the expression of HOTTIP can significantly reduce the proliferation ability of HSC-4 cells.In the cell scratch test,the scratch healing speed of the cells in the HOTTIP-inhibitor group was slowed down.The percentage of cell scratches(53.27±1.85)in the HOTTIPinhibitor group at 24 h after transfection was greater than(21.10.±0.78)and(25.13±1.69)in the Control group and Inhibitor-NC group,and the differences were statistically significant(P<0.05),suggesting that the low expression of HOTTIP can effectively reduce the cell migration ability.Conclusion Compared with HOK cells,the expression of HOTTIP in HSC-4 cells was significantly up-regulated.Knocking down the expression of HOTTIP can significantly reduce the proliferation and migration of HSC-4 cells,confirming that it plays a pro-cancer role in OSCC.