Objective To evaluate the biological effects of human platelet-derived growth factor-B gene transfection on gingival fibroblasts . Methods The plasmid(EX-A0380-M03) carried human plate-let-derived growth factor-B(hPDGF-B) gene was transfected into gingival fibroblasts by LipofectamineTM 2000 . Morphological changes of transfected cells were recorded by optical and fluorescent microscope . Then ,MTT and flow cytometry were used to estimate the proliferation and apoptosis of gene-modified cells . The expression of collagen I and XII ,as well as theα-SMA(α-Smooth Muscle Actin) was detected by Real-time Quantitative PCR . Results Compared to the normal cells ,partial transfected cells presen-ted startlike ,polygonal ,and neuron-like . Cells proliferation was enhanced after gene interference ,but apoptosis did not appeared . At transcription level ,the expression of Collagen I was up-regulated ,while Collagen XII and α-SMA were down-regulated . Conclusion The enhanced proliferation of gingival fibro-blasts were due to paracrine and autocrine modes of fusion protein PDGF-BB-eGFP synthetized by gene modified cells . Transfeced cells did not present reverse differentiation ,and the extracellular matrix re-modeling ability of those cells were down . The synthesis of Collagen I of hPDGF-B gene modified cells ascended no tably .%目的：检测hPDGF-B基因转染对Beagle犬牙龈成纤维细胞生物学活性的影响。方法使用脂质体LipofectamineTM 2000将携带hPDGF-B基因的质粒EX-A0380-M03转染至Beagle犬牙龈成纤维细胞，观察转染后细胞的形态以及增殖的变化；流式细胞仪检测转染细胞的周期以及凋亡情况；Real-time PCR法检测转染后细胞表达Collagen I、Collagen XII、α-SM A的变化。结果转染后的牙龈成纤维细胞形态多样，但增殖显著；转染组S期细胞比率增高，转染细胞未出现凋亡；转染后细胞上调Collagen I基因的表达，下调Collagen XII和α-SM A的表达。结论 hPDGF-B基因转染牙龈成纤维细胞后，产生PDGF-BB-eGFP融合蛋白，融合蛋白可能通过自分泌和旁分泌的方式作用于牙龈成纤维细胞，从而促进细胞增殖。hPDGF-B基因转染牙龈成纤维细胞后，细胞并不能出现逆向分化，细胞对外周基质的改建能力下降，但细胞合成胶原的能力显著增强。
展开▼
