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目的探讨莱菔硫烷(sulforaphane,SF)对鼻咽癌细胞中Krüppel样因子4(Krüppel-like factor 4,Klf4)表达及侵袭、转移能力的影响。方法将鼻咽癌细胞株HONE1和5-8F分为HONE1对照组、HONE1SF处理组、5-8F对照组、5-8FSF处理组,HONE1对照组和5-8F对照组正常培养,不做任何处理;HONE1SF处理组和5-8FSF处理组细胞采用梯度浓度SF处理。72 h后采用CCK-8实验检测细胞存活率,并计算半数抑制浓度(half maximal inhibitory concentration,IC_(50));取50%IC_(50)浓度SF处理HONE1和5-8F后,采用免疫荧光法观察Klf4的亚细胞定位,采用划痕试验和Transwell试验观察细胞侵袭迁移能力,采用Western blot法检测E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(vimentin)等上皮-间充质细胞转化相关蛋白表达情况。结果经0~20.0μmol/L SF处理后的HONE1和5-8F细胞存活率逐渐下降,SF的IC_(50)值分别为8.6和9.4μmol/L;HONE1SF处理组和5-8FSF处理组细胞质中Klf4表达明显高于HONE1对照组和5-8F对照组,细胞迁移速度低于HONE1对照组和5-8F对照组,侵袭细胞数少于HONE1对照组和5-8F对照组;HONE1SF处理组E-cadherin表达量[(17 543 490±7 545)Int]高于HONE1对照组[(13 543 656±4 554)Int],N-cadherin[(14 239 989±6 902)Int]及vimentin[(6 090 321±3 475)Int]低于HONE1对照组[(16 093 409±9 348)、(8 994 382±8 543)Int](P<0.05);5-8FSF处理组E-cadherin表达量[(15 430 038±6 821)Int]高于5-8F对照组[(12 397 373±6 432)Int],N-cadherin[(11 980 109±2 343)Int]及vimentin表达量[(4 903 480±3 090)Int]低于5-8F对照组[(15 999 890±2 390)、(7 098 023±5 409)Int],差异均有统计学意义(P<0.05)。结论 SF可导致鼻咽癌细胞Klf4在细胞质中聚集,降低其在体外的侵袭迁移能力,可能与细胞质中的Klf4升高E-cadherin表达、降低vimentin表达有关。
Objective To investigate the effect of sulforaphane (SF) on the expression and invasion and metastasis of Krüppel-like factor 4 (Klf4) in nasopharyngeal carcinoma cells. Methods The nasopharyngeal carcinoma cell lines HONE1 and 5-8F were divided into HONE1 control group, HONE1SF treatment group, 5-8F control group, 5-8FSF treatment group, HONE1 control group and 5-8F control group without any treatment ; HONE1SF treatment group and 5-8FSF treatment group cells treated with gradient concentration of SF. After 72 h, the cell viability was measured by CCK-8 assay and the half maximal inhibitory concentration (IC 50) was calculated. After treatment of HONE 1 and 5-8 F with 50% IC 50, SF was detected by immunofluorescence Method was used to observe the subcellular localization of Klf4. The invasion and migration of cells were observed by scratch test and Transwell assay. The expression of E-cadherin, N-cadherin, vimentin (vimentin) and other epithelial-mesenchymal transition related protein expression. Results The survival rates of HONE1 and 5-8F cells treated with 0 ~ 20.0μmol / L SF decreased gradually, and the IC 50 values of SF were 8.6 and 9.4μmol / L, respectively. In the cytoplasm of HONE1SF and 5-8FSF treated groups, Klf4 expression was significantly higher than the HONE1 control group and 5-8F control group, the cell migration rate was lower than the HONE1 control group and 5-8F control group, the number of invasive cells less than the HONE1 control group and 5-8F control group; HONE1SF treatment group E- (17 543 490 ± 7 545) Int] was higher than that in HONE1 control group [(13 543 656 ± 4 554) Int], N-cadherin [(14 239 989 ± 6 902) Int] and vimentin [(6 090 321 ± 3 475) Int] was lower than that in HONE1 control group [(16 093 409 ± 9 348), (8 994 382 ± 8 543) Int] (P <0.05) (15 430 038 ± 6 821) Int] was higher than that in 5-8F control group [(12 397 373 ± 6 432) Int], N-cadherin [(11 980 109 ± 2 343) Int] 903 480 ± 3 090) Int] was lower than that of 5-8F control group [(15 999 890 ± 2 390), (7 098 023 ± 5 409) Int], the differences were statistically significant (P <0.05). Conclusion SF can induce the accumulation of Klf4 in the cytoplasm of nasopharyngeal carcinoma cells and decrease its invasion and migration in vitro. It may be related to the increase of E-cadherin expression and the decrease of vimentin expression by Klf4 in cytoplasm.