CAJ | 학술논문

目的：体外实验研究PLCε基因调节人膀胱癌细胞迁移和侵袭的分子机制。方法设计并合成针对PLCε基因mRNA的寡核苷酸序列，构建重组腺病毒Ad-shPLCε。 T24细胞感染Ad-shPLCε腺病毒后，用Western blot检测PKCα／β及TBX3、E-cadherin的表达；用划痕实验、Transwell 迁移和侵袭实验检测膀胱癌细胞T24的迁移、侵袭能力。结果 Ad-shPLCε重组腺病毒感染膀胱癌细胞T24，对其PLCε mRNA表达抑制率为75.6％，对其PLCε蛋白表达抑制率为67.4％。 Ad-shPLCε感染组T24细胞迁移能力较空载组和空白对照组明显减弱（ P＜0.05）；重组腺病毒Ad-shPLCε感染组T24细胞穿膜细胞数较空载组和空白对照组明显减少（ P＜0.05）。 Ad-shPLCε重组腺病毒感染膀胱癌T24细胞后下游PKCα／β的活化受到抑制（ P＜0.05），同时，TBX3的表达减少，而E-cadherin的表达增高（ P＜0.05）。结论通过以重组腺病毒干扰抑制PLCε基因，能有效抑制膀胱癌T24细胞系中PLCε下游PKCα／β的活化情况及TBX3，E-cadherin的表达变化，并且对T24细胞的迁移、侵袭能力具有一定的抑制作用。
목적：체외실험연구PLCε기인조절인방광암세포천이화침습적분자궤제。방법설계병합성침대PLCε기인mRNA적과핵감산서렬，구건중조선병독Ad-shPLCε。 T24세포감염Ad-shPLCε선병독후，용Western blot검측PKCα／β급TBX3、E-cadherin적표체；용화흔실험、Transwell 천이화침습실험검측방광암세포T24적천이、침습능력。결과 Ad-shPLCε중조선병독감염방광암세포T24，대기PLCε mRNA표체억제솔위75.6％，대기PLCε단백표체억제솔위67.4％。 Ad-shPLCε감염조T24세포천이능력교공재조화공백대조조명현감약（ P＜0.05）；중조선병독Ad-shPLCε감염조T24세포천막세포수교공재조화공백대조조명현감소（ P＜0.05）。 Ad-shPLCε중조선병독감염방광암T24세포후하유PKCα／β적활화수도억제（ P＜0.05），동시，TBX3적표체감소，이E-cadherin적표체증고（ P＜0.05）。결론통과이중조선병독간우억제PLCε기인，능유효억제방광암T24세포계중PLCε하유PKCα／β적활화정황급TBX3，E-cadherin적표체변화，병차대T24세포적천이、침습능력구유일정적억제작용。
Objective To investigate the molecular mechanisms of PLCεin regulating the invasion and migration of human bladder cancer cells in vitro.Methods After cells treated with recombinant adenovirus , the migratory/in-vasive abilities of T24 cells were explored by wound healing and Transwell chamber cell migration and invasion as -say;RT-PCR was used to detect the mRNA levels of PLCε;The protein levels of PLCε,PKCα,PKCβ, TBX3 and E-cadherin were determined by Western blot;QRT-PCR was used to detect the mRNA levels of TBX3 and E-cad-herin.Results It was confirmed by digesting and sequencing that the recombinant adenovirus had been constructed successfully .The expression of PLCε mRNA and PLCε protein were both decreased after the infection of Ad-shPLCε.Wound healing and Transwell chamber cell migration/invasion assay showed that Ad-shPLCε treatment could inhibit the migratory and invasive activity of bladder cancer cells(P<0.05).The results of Western blot indicated that the expression of PKCα/βin membrane decreased ( P<0.05 ) , and phosphorylation level of PKCαand PKCβwas reduced .QRT-PCR and Western blot analysis demonstrated that the expression level of TBX 3 de-creased , but the expression level of E-cadherin increased .Conclusions PLCε shRNA can inhibit migratory and invasive ability of bladder cancer cells through PKCα/β/TBX3/E-cadherin pathway .