| Objective:Tongue squamous cell carcinoma is one of the most common malignant tumor in oral and maxillofacial region, which has a high mortality rate. Due to the rich lymphatic and blood supply, as well as the frequent mechanical movement of the tongue, tumor cell metastasize by blood and lymphatic vessel easily. At present, operation is still the preferred, but the dysfunction and appearance deformity caused by surgery is hard to bearing. In recent years, an increasing number of reports showed that potassium ion channels, especially voltage-gated potassium ion channel, are involved in a variety of physiological activities of cells. It is abnormality expressed in many tumor cells, but have low expression in the correspongding normal tissues. The voltage-gated potassium ion channels are involved in tumor cell proliferation, differentiation and signal processes of tumor formation, participating in tumor development.4-aminopyridine(4-AP) is one of the potassium channel blocker, which can selectively blocking Kv channels. Our previous study found that 4-AP can not only inhibited the proliferation of human tongue squamous cell carcinoma Tca8113 cells but also induce or promote Tca8113 cell apoptosis. To explore the effect of 4-AP on the migration and invasion of Tca8113 cell, thus provide reference for clinical studies. Methods:Human tongue squamous cell carcinoma Tca8113 was subcultured, which in exponential phase of growth was chosen to be treated as followed.1. Wound healing assay was employed to explore whether 4-AP (10,20,30mmol/L)could inhibit the migration of Tca8113 cell.2. Transwell assay was carried out to detect the effect of 4-AP on the invasion of Tca8113 cell.3. All data were analyzed by SPSS 17.0 software, and they expressed by x=s. The LSD test and Tamhane test were used for cell migration rate and Transwell assay post hoc multiple comparisons respectively. A value of P<0.05 was considered to be significant. Results:1. The inverted phase contrast microscope shows that, the Tca8113 cells did not change significantly after adding 10,20 and 30 mmol/L 4-AP for 24 hours and the cell growing in a good condition. While, with the increasing concentration of 4-AP and culture time, the cell shrinked and turned round, and the volume of cells become decreased, cells occurred a series of morphological changes.2. The potassium channel blocker 4-AP can inhibited the human tongue squamous cell carcinoma cell Tca8113 cells. The scratch healing assay obtained cell migration rates of (19.25±4.50)%, (14.07±5.59)%, (11.34±1.79)% and (8.71±3.26)% for Tca8113 cells treated with 0,10,20,30mmol/L of 4-AP for 12h and (33.16±6.40)%, (22.20±5.30)%, (14.24±2.52)% and (11.65±2.97)% for 24h. The results showed that the migration rate were significantly difference between 4-AP goups(10,20,30mmol/L)and control groups(P<0.01). Moreover, the scratch healing assay result indicated that 4-AP could suppress the migration of Tca8113 cells in a dose-dependent manner. There was no significantly difference between 4-AP goups 20mmol/L and 10mmol/L in 12h, 20mmol/L and 30mmol/L in 12h and 24h(P>0,05). The rest of the experimental group were significant difference(P<0.05).3.The transwell invasion assay showed that with the increasing of 4-AP concentration, the Tca8113 cell number decrased. The number of cells permeating through the matrigel was(112.20±24.36), (28.87±6.47), (11.80±5.77) when the Tca8113 cells were treated with 10mmol/L,20mmol/L,30mmol/L 4-AP for 24h, while the control group was (179.73±16.14), and the difference was significant(F=392.045, P<0.001). Conclusion:4-AP can significantly inhibited the migration and invasion of human tongue carcinoma cell line Tca8113. |
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