c—myc靶向小干擾RNA誘導乳腺癌細胞

翟榮林王國斌 夏澤峰 【摘要】 耳的構建小干擾RNA(siRNA)表達載體，體外觀察其誘導乳腺癌細胞凋亡效應。 方法基因克隆技術構建人c—mye癌基因特異性siRNA表達載體，體外轉染MCF．7。48 h后檢測c—my茈基因表達狀況并觀察MCF一7凋亡誘導效應。結果 (1)成功構建siRNA表體載體：pEGFP—C1／U6．1、pEGFP—c1／U6—2和pEGFP—C1／U6．3；(2)siRNA表達載體轉染MCF一7 48 h后。pEGFP-C1／U6．2組顯著抑制胞內c．myc表達(80％)；(3)轉染24 h和48 h后，pEGFP．C1／U6—2組凋亡率分別為11．01％和21．30％。明顯高于對照組(P<0．01)。結論構建的c—myc靶向siRNA表達載體能顯著下調c．myc在MCF-7中的過表達。誘導乳腺癌細胞株MCF-7凋亡。 【關鍵詞】 c．myc基因； siRNA； 乳腺癌； 脫噬作用 Apoptasis induction in breast~ancef cells by siRNA targeting gene c-myc 【Abstract】 Objective To observe apoptoais induction in breast cancer cells by siRNA．Methods The siRNA expression vectors were constructed and transfected into MCF一7．At 48 h after transfection． cell apoptosis in MCF-7 was detected by FCM ．The gene expression of C-ITIyc was detected bv RT．PCR and Western blotting respectivdy．Results (1)Transfectbn of pEGFP—C1／U6—2 group into MCF-7 down-regulated C-myc expression level greatly at 48 h after transfection(80％)；(2)The apoptosis per—centage of pEGFP—C1／U6．2 group was 11．01％ at 24 h and 21．30％ respectivdy at 48 h after transfec． tion。significantly higher than in the control group(P<0．01)．Conclusion The siRNA expression vec—tor we comt~ct can effectively down-regulate c-myc over expression and remarkably induce apoptosis in MCF一7 cell line． 【Key words】c-myc gene； siRNA； Breast carcinoma； Apoptosis

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