發(fā)布時(shí)間：2019-05-29 01:20

【摘要】：目的探討過(guò)表達(dá)脾酪氨酸激酶(SYK)對(duì)結(jié)直腸癌細(xì)胞增殖和凋亡的影響及可能的相關(guān)機(jī)制。方法利用pcDNA.3.1質(zhì)粒構(gòu)建重組質(zhì)粒pcDNA.3.1-SYK,轉(zhuǎn)染結(jié)直腸癌細(xì)胞,過(guò)表達(dá)SYK,分組情況如下。(1)pcDNA.3.1-SYK(HCT116):轉(zhuǎn)染pcDNA.3.1-SYK到HCT116;(2)pcDNA.3.1(HCT116):轉(zhuǎn)染pcDNA.3.1空載體到HCT116中;(3)Normal(HCT116):正常HCT116細(xì)胞。(1)pcDNA.3.1-SYK(Sw480):轉(zhuǎn)染pcDNA.3.1-SYK到Sw480;(2)pcDNA.3.1(Sw480):轉(zhuǎn)染pcDNA.3.1空載體到Sw480中;(3)Normal(Sw480):正常Sw480細(xì)胞。應(yīng)用q RT-PCR法檢測(cè)結(jié)直腸癌和癌旁組織中SYK和Fra-1的mRNA表達(dá)量;Western blot法檢測(cè)SYK和Fra-1的蛋白表達(dá)量;MTT法檢測(cè)細(xì)胞生長(zhǎng)活力;Brd U方法檢測(cè)細(xì)胞增殖活性;試劑盒方法檢測(cè)Caspase-3的活性;Annexin-V FITC/PI法檢測(cè)細(xì)胞凋亡情況。結(jié)果 SYK在結(jié)直腸癌組織和結(jié)直腸癌細(xì)胞系中的表達(dá)量均降低(P0.01);pcDNA.3.1-SYK轉(zhuǎn)染結(jié)直腸癌細(xì)胞系,SYK的mRNA(P0.01)和蛋白表達(dá)量顯著升高(P0.01),顯示SYK過(guò)表達(dá)成功;SYK過(guò)表達(dá)后結(jié)直腸癌細(xì)胞生長(zhǎng)活力和增值活性顯著降低(P0.01),細(xì)胞凋亡增加(P0.01);另外,SYK過(guò)表達(dá)后,Fra-1的表達(dá)量顯著被抑制(P0.01)。結(jié)論過(guò)表達(dá)SYK對(duì)結(jié)直腸癌細(xì)胞的增殖有抑制作用,并且促進(jìn)結(jié)直腸癌細(xì)胞的凋亡,其機(jī)制有可能與SYK對(duì)Fra-1的調(diào)控有關(guān),為結(jié)直腸癌的預(yù)防和治療提供參考價(jià)值和理論基礎(chǔ)。
[Abstract]:Objective to investigate the effect of overexpression of spleen tyrosine kinase (SYK) on proliferation and apoptosis of colorectal cancer cells and its possible mechanism. Methods the recombinant plasmid pcDNA.3.1-SYK, was constructed by pcDNA.3.1 plasmid and transfected into colorectal cancer cells. The overexpression of SYK, was divided into the following groups. (1) pcDNA.3.1-SYK (HCT116): pcDNA.3.1-SYK into HCT116; (2) pcDNA.3.1 (HCT116): transfer pcDNA.3.1 empty vector into HCT116; (3) Normal (HCT116): normal HCT116 cells. (1) pcDNA.3.1-SYK (Sw480): pcDNA.3.1-SYK to Sw480; (2) pcDNA.3.1 (Sw480): pcDNA.3.1 empty vector was transferred into Sw480, (3) Normal (Sw480): normal Sw480 cells. The mRNA expression of SYK and Fra-1 in colorectal cancer and paracancerous tissues was detected by Q RT-PCR assay. The protein expression of SYK and Fra-1 was detected by; Western blot assay, and the cell proliferation activity was detected by MTT assay.; Brd U assay was used to detect the proliferation activity of colorectal cancer and paracancerous tissues. The activity of Caspase-3 was detected by kit method and apoptosis was detected by Annexin-V FITC/PI assay. Results the expression of SYK in colorectal cancer tissue and colorectal cancer cell line decreased (P 0.01). The mRNA (P01) and protein expression of SYK were significantly increased (P01) in colorectal cancer cell line pcDNA.3.1-SYK, which showed that the overexpression of SYK was successful. After overexpression of SYK, the growth activity and value-added activity of colorectal cancer cells decreased significantly (P01), and apoptosis increased (P01). In addition, after overexpression of SYK, the expression of Fra-1 was significantly inhibited (P01). Conclusion overexpression of SYK can inhibit the proliferation of colorectal cancer cells and promote the apoptosis of colorectal cancer cells. The mechanism may be related to the regulation of Fra-1 by SYK, which provides a reference value and theoretical basis for the prevention and treatment of colorectal cancer.
【作者單位】： 河南省中醫(yī)院肛腸科;河南中醫(yī)藥大學(xué)中醫(yī)外科;
【基金】：河南省中醫(yī)藥科學(xué)研究專項(xiàng)課題項(xiàng)目(2015ZY02049)
【分類號(hào)】：R735.3

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