Sự đảo ngược hoàn toàn quá trình chuyển đổi biểu mô sang trung mô yêu cầu ức chế cả sự biểu hiện của ZEB và con đường Rho
Tóm tắt
Quá trình chuyển đổi biểu mô sang trung mô (EMT) liên quan đến yếu tố tăng trưởng biến đổi-β (TGF-β) là một sự kiện tế bào quan trọng trong quá trình hình thành cơ quan, ung thư và xơ hóa cơ quan. Quy trình đảo ngược EMT vẫn chưa được thiết lập rõ ràng. Mục đích của chúng tôi là xác định các con đường truyền tín hiệu và các yếu tố phiên mã duy trì trạng thái trung mô do TGF-β gây ra.
Các chất ức chế năm loại kinase có liên quan đến EMT, bao gồm kinase thụ thể TGF-β loại I (TβRI), kinase p38 MAPK, kinase kinase MAP/extracellular signal-regulated kinase activator kinase (MEK1), kinase c-Jun NH-terminal (JNK), và kinase Rho (ROCK), đã được đánh giá về khả năng đảo ngược trạng thái trung mô được gây ra trong các tế bào biểu mô ống thận. Các tác nhân đơn lẻ không hoàn toàn đảo ngược EMT như được xác định bởi hình thái tế bào và biểu hiện gen. Tuy nhiên, việc tiếp xúc với chất ức chế TβRI SB431542, kết hợp với chất ức chế ROCK Y27632, đã loại bỏ các sợi căng stress actin có thể phát hiện và biểu hiện gen trung mô trong khi phục hồi biểu hiện E-cadherin và cadherin đặc hiệu thận (Ksp-cadherin). Một sự kết hợp thứ hai, chất ức chế TβRI SB431542 cùng với chất ức chế p38 MAPK SB203580, cũng có tác dụng một phần trong việc đảo ngược EMT. Hơn nữa, chất ức chế JNK SP600125 ức chế hiệu quả của chất ức chế TβRI SB431542 trong việc đảo ngược EMT. Để khám phá cơ sở phân tử dưới đây đảo ngược EMT, chúng tôi cũng đã nhắm đến các yếu tố ức chế phiên mã ZEB1 và ZEB2/SIP1. Giảm biểu hiện ZEB1 và ZEB2 trong các tế bào tuyến vú của chuột bằng shRNAs đã đủ để tăng cường biểu hiện của các protein biểu mô như E-cadherin và khôi phục các đặc điểm biểu mô. Tuy nhiên, việc phục hồi hoàn toàn cortical F-actin cần phải ủ với chất ức chế ROCK Y27632 kết hợp với sự gõ của ZEB1/2.
Từ khóa
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