Abstract
Cisplatin is one of the major drugs used in oral cancer treatments, but its usage can be limited by acquired drug resistance. In this study, we established three cisplatin-resistant oral squamous cell carcinoma (OSCC) cell lines and characterized them using cell viability assays, qPCR, Western blotting, FACS, immunofluorescence, and wound healing assays. Three OSCC cell lines (YD-8, YD-9, and YD-38) underwent long-term exposure to cisplatin, eventually acquiring resistance to the drug, which was confirmed by an MTT assay. In these three newly established cell lines (YD-8/CIS, YD-9/CIS, and YD-38/CIS), overexpression of multidrug resistance (MDR)-related genes was detected by qPCR and Western blotting. The cell lines displayed an increase in the functional activities of breast cancer resistance protein (BCRP) and multidrug resistance protein1 (MDR1) by rhodamine 123 and bodipy FL prazosin accumulation assays. Moreover, the cisplatin-resistant cells underwent morphological changes, from round to spindle-shaped, increased expression of epithelial-to-mesenchymal transition (EMT)-related molecules such as N-cadherin, and showed increased cell migration when compared with the parental cell lines. These results suggest that these newly established cell lines have acquired drug resistance and EMT induction.
Highlights
Oral cancer is an increasingly global disease [1,2], more than half of these cancers are found late, sometimes permanently altering a patient’s ability to chew, swallow, talk, as well as their appearance [3]
Parental and cisplatin-resistant oral squamous cell carcinoma (OSCC) cell lines. (a) The expression of multidrug resistance (MDR)-related genes was measured by qPCR. * p < 0.01 versus parental cells. (b) Left, the expression of multidrug resistance protein1 (MDR1) and breast cancer resistance protein (BCRP) proteins was determined by a Western blot assay
We examined MDR1 protein activity using rhodamine 123, and BCRP protein activity using bb9o/oCddIiiSppWyyceeFFlaLlLlsspop(r3rae4azx.zo4ao%smsin)iinn.c.eFoFdimgigMupuraDerree3Rda31astphosrohopwotaewsrinetshnatatchattaltivhtcieetthlyalescuca(su5cimn4cu.gu2m%lrahu)to.ilodaInntaimootnfhinreoheftoh1drr2hae3ome,daicannimsedp1ilBn2aCe3tiRnw12P-ar3espswrrieosadttseauinrcnteedadccueitlncilveYliditnDyie-nus9,s/YCitnDhIgSe1caiYrnee7cDltsc.l5ruus-a%8lm(ct/3s;eC4ulYI.lsl4uSDah%,ltoa-1i8)owr7,cn.ao56ctom%5chf.u0ap;bmYt%aorDYu;deY-dlDia8pDt-t,yoi86-o,95pFn/.YaL0CorD%eIfpSn-;br9,tYaoa,2zDdl5aoci.-np8se9id%ly/nlCs;FYwI(YL5SDa4,Dps.-223-r5l9%a8e.,8zs)6%co.s6esIt;l.nih0lYns%atDhwnd;e-ae9Yitnsv,hD6elrte6le-hos3e.e0sp8ci%/etriChsd;ppaIYaSlncaDr,iteis5i-nnpn32tl-8t.aa0rh/etl%CeiscniIi;resS-Ytrlp,ale5Dansl2riit-ne.s30ctne8e%ast,lnal;7(cllFY8iecni.Deg2eblu%-sly3,rl8)tie.hn,in37eTecb8sihrn.;(2eetFYas%rieasDg)ic.run-eeT8rgls/lehuCute3llhIastbSeser;, sahctoiwvittyhoatf MYDD-R8,1YoDr B-9C, RanPd
Summary
Oral cancer is an increasingly global disease [1,2], more than half of these cancers are found late, sometimes permanently altering a patient’s ability to chew, swallow, talk, as well as their appearance [3]. It is important to treat oral cancers at the early stages with minor surgery. Overall the treatment requires an efficient combination of surgery, radiotherapy, and chemotherapy [4,5]. A platinum-based drug, is one of the standard first-line chemotherapeutic agents used to treat OSCC [6]. Several factors can lead to the failure of chemotherapy treatments, including acquired resistance to the chemotherapeutic agents [7,8]. Some research has shown that acquired chemoresistance is associated with overexpression of the ATP-binding cassette (ABC) transporters, which pump drugs out of cells [9,10]. There are 48 known ABC transporters in humans, including MRP1-6, MRP9, MDR1, and BCRP [10,11]
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