The Effect Of Voltage-gated Sodium Channels Nav1.5-E3 Antibody On Migration And Invasion Of SiHa, MDA-MB-231 And Caov-3 Cells

To explore the inhibition effects of a subtype-specific inhibitor of NaV1.5 by E3 targeting(Nav1.5 - E3 antibody) in SiHa cervical cancer cell, MDA-MB-231 breast cancer cell andCaov-3 ovarian cancer cell in vitro and in vivo.Keywords Nav1.5 - E3 antibody; Breast cancer; ovarian cancer; transplantable tumorMethods1. Use Real-time PCR to analyze the expression of Nav1.5mRNA.2. Transwell chamber migration assay was used to detect the migrating ability of cells invitro.3. Transwell chamber invasion assay was used to detect the invasion capacity of cells invitro.4. Human cervical cancer SiHa, breast cancer MDA-MB-231, and ovarian cancer Caov-3cells were subcutaneously transplanted into nude mice to establish xenograft tumors. UseNav1.5 - E3 antibody, liducaine and PBS to interfere in the three cells,every group had10 nude mice(n=10). The tumor volume was measured and the tumor morphology wasobserved by HE staining.Results1. RT-PCR showed that Nav1.5mRNA level were significantly decreased in cells exposedto Nav1.5-E3 or lidocaine BPS P<0.001 , and the difference between Nav1.5-E3 andlidocaine groups was significant P<0.05 . 2. Migration assays have showed that 20ug/mLNav1.5 - E3 antibody or 200ug/mLlidocaine could reduce the number of SiHa, MDA - MB - 231 and Caov - 3 cells whichacrossed through the membrane of the Transwell chamber. Count the number of the cellsand compared respectively showed the difference with a statistical significance, P <0.001.But lidocaine group and Nav1.5 E3 antibody group there were no significant differencesin every cell.3. The invasion assay showed that Nav1.5- E3 antibodies and lidocaine affected theinvasion ability of SiHa, MDA - MB-231 and Caov - 3 cells. Compared with theirindividual control group, the Nav1.5- E3 antibodies and lidocaine droped the cells whichcould traverse the the membrane of the Transwell chamber, the difference were statisticalsignificant (P<0.001). But between the lidocaine group and Nav1.5-E3 antibody groupno significant differences were found.4. In nude mice model, after using antibody and lidocaine, the tumor volume wassignificantly decreased in Nav1.5-E3 antibody groups and liducaine groups, comparedwith that of the PBS control group. The inhibition rate of E-3 antibody, lidocaine were32.18% and 37.61%, 28.59% and 39.12%, 28.53% and 30.77% in SiHa, MDA-MB-231,and Caov-3 cells respectively. The discrepancy was significant P<0.01 .Themorphology is definite changes in tumors E-3 antibody and lidocaine groups and PBScontrol groups. The necrosis and apoptosis of tumor cells were found under lightmicroscope more frequent in Nav1.5 antibody and lidocaine groups than in PBS groups.ConclusionNav1.5-E3 antibody targeting antibody can depress the expression of Nav1.5 in cancercells. Nav1.5-E3 antibody can inhibit the transplantable tumor of the human cervicalcancer SiHa, breast cancer MDA-MB-231 and ovarian cancer Caov-3. Nav1.5-E3antibody may play an important role in therapy for cancers. Nav1.5 - E3 antibody cansignificantly reduce the growth of transplant tumors. This effect may be due to theinhibition of the expression of the iron channel in cells.