CLIC1 Participates In The Migration And Invasion Of SiHa Cells By Regulating Intracellular ROS

Objective:Owing to the high incidence and metastasis rate,cervical cancer has always occupied a significant position in female malignant tumors,therefore,it is of extraordinary significance to clarify the mechanism of metastasis and diffusion of cervical cancer cells for its clinical strategy exploration.The role of intracellular chloride channel protein1(CLIC1)in the progression of malignant tumors has been confirmed by several studies at home and abroad,but the specific functions and mechanisms remain unclear.The malformed vascular system is a common feature of most malignant tumors.This abnormal vascular system causes chronic changes in the growth environment around tumor cells,leading to a long-term sustained hypoxia-reoxygenation(H-R)environment.And the most direct result of this shift is the increase in intracellular reactive oxygen species(ROS).Moreover,the malignant phenotype of tumor cells stimulated by ROS accelerates the malignant progression of tumor furtherly.This study observed whether CLIC1 under H-R conditions could promote the progression of cervical cancer by altering intracellular ROS levels.Methods:Human SiHa cell lines were selected for subculture,and the H-R model and normoxic model were constructed by physical conditions.SiHa cells in different culture environments were treated with IAA94(20?mol/l),DPI(15?mol/l)and NAC(20 mmol/l).Relative expression of CLIC1 protein in each group was detected by Western-Blot;Detection of intracellular ROS content uses DCFH-DA fluorescent probe kit;Scratch and cell invasion experiments are used to observe changes in migration and invasion of SiHa cells under normal condition and H-R Condition.SPSS 22.0 software is needed for final processing analysis of acquired data.Results:1.There is a remarkble ascending trend for the relative expression of CLIC1 protein in the H-R group(3.57±0.03)when comparing to the normoxia group(1.00±1.18)(P<0.01),and the relative expression of CLIC1 protein in the H-R+IAA94 group(2.39±0.01)is lower in comparison with cells cultured in the hypoxia and reoxygenation environment(3.57 ±0.03)(P<0.01).The relative expression of CLIC1 protein in the H-R+DPI group and the H-R+NAC group(2.83±0.03,2.85±0.02)show a descending trend versus the H-R group(3.57±0.03)(P<0.01).2.The relative content of ROS in H-R group acts an obvious rising trend versus normoxia group(P<0.01),while the relative content of ROS in H-R+IAA94 group was lower than that in H-R group(P<0.01).The relative content of ROS in H-R+DPI group and H-R+NAC group were also lower than that in H-R group(P<0.01).3.The scratch healing area(61.76±0.52)of SiHa cells in the H-R group is dramatically larger than those SiHa cells cultured in the enviroment full of oxygen(41.94±1.18)(P<0.01),while the scratch healing area(50.53±3.1)in the H-R+IAA94 group is smaller in comparison with the H-R ones(61.76±0.52)(P<0.01).The scratch healing area of the H-R+DPI group and H-R+NAC group(49.34±2.15,49.21±3.13)are both smaller versus the H-R group(61.76±0.52)as well(P<0.01).4.In comparison with cells in hypoxia and reoxygenation(91.80±10.96),the number of invasive SiHa cells in normal condition(46.60±6.19)is decreasing(P<0.01),what's more,there is also an obvious downtrend in the the number of SiHa cells arriving the lower layer after treated with IAA94(69.20±9.88)(P<0.01),and the same to the SiHa cells in the H-R+DPI group and the H-R+NAC group(65.20±9.58,64.40±6.80)(P<0.01).Conclusion:CLIC1 promotes cervical cancer progression by regulating intracellular ROS to involve in the migration and invasion of cervical cancer SiHa cells in the hypoxic-reoxygenation environment.Therefore,clinical strategies using CLIC1 and ROS as relevant pharmacological targets may provide hope for cervical cancer treatment.