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Abstract：
BACKGROUND: The pathogenesis of fibrous epulis is still quite unclear. Our recent genome-wide RNA sequencing analysis revealed that in fibrous epulis, RAS-PI3K-AKT-NF-κB pathway regulates the expression of Bcl-2 family and IAP family genes, leading to increased proliferation and the inhibition of apoptosis. The PI3K/AKT signaling pathway can promote autophagy in human gingival fibroblasts; therefore, the purpose of the present study was to identify whether autophagy is involved in the pathogenesis of fibrous epulis.
METHODS: Differentially expressed genes (DEGs) between fibrous epulis lesions and normal gingival tissues were identified using the PCR array. The expression levels of eighteen autophagy-related (ATG) family genes, twelve B-cell lymphoma 2 (Bcl-2) family genes, and eleven cysteine-dependent aspartate-directed protease (caspase) family genes were validated using quantitative real-time PCR (qRT-PCR). Autophagy induction was determined by measuring microtubule-associated protein light chain 3 (LC3) conversion (LC3-I to LC3-II) by immunoblot analysis.
RESULTS: The PCR array identified six upregulated genes, whereas no genes were expressed at significantly lower levels. The upregulated genes were BCL2, BCL2L1, CXCR4, HSP90AA1, HSPA8, and IGF1, which all belong to the \"regulation of autophagy\" group but not the \"autophagy machinery components\" group. qRT-PCR verified that the expression levels of BCL2, BCL2L1 (also known as BCL-XL), and BCL2L2 (also known as BCL-W) were significantly increased in fibrous epulis. No LC3-I to LC3-II conversion was observed.
CONCLUSIONS: The present study reveals that in fibrous epulis, Bcl-2 and Bcl-xL coordinately mediate gingival cell escape from apoptosis, leading to uncontrolled proliferation. Moreover, ATG family genes are not activated, and autophagy is not involved in this process.
METHODS: Differentially expressed genes (DEGs) between fibrous epulis lesions and normal gingival tissues were identified using the PCR array. The expression levels of eighteen autophagy-related (ATG) family genes, twelve B-cell lymphoma 2 (Bcl-2) family genes, and eleven cysteine-dependent aspartate-directed protease (caspase) family genes were validated using quantitative real-time PCR (qRT-PCR). Autophagy induction was determined by measuring microtubule-associated protein light chain 3 (LC3) conversion (LC3-I to LC3-II) by immunoblot analysis.
RESULTS: The PCR array identified six upregulated genes, whereas no genes were expressed at significantly lower levels. The upregulated genes were BCL2, BCL2L1, CXCR4, HSP90AA1, HSPA8, and IGF1, which all belong to the \"regulation of autophagy\" group but not the \"autophagy machinery components\" group. qRT-PCR verified that the expression levels of BCL2, BCL2L1 (also known as BCL-XL), and BCL2L2 (also known as BCL-W) were significantly increased in fibrous epulis. No LC3-I to LC3-II conversion was observed.
CONCLUSIONS: The present study reveals that in fibrous epulis, Bcl-2 and Bcl-xL coordinately mediate gingival cell escape from apoptosis, leading to uncontrolled proliferation. Moreover, ATG family genes are not activated, and autophagy is not involved in this process.
摘要：
背景：纤维性上皮的发病机制尚不清楚。我们最近的全基因组RNA测序分析显示，RAS-PI3K-AKT-NF-κB通路调控Bcl-2家族和IAP家族基因的表达,导致增殖增加和凋亡抑制。PI3K/AKT信号通路可促进人牙龈成纤维细胞的自噬;本研究的目的是确定自噬是否参与纤维性血管的发病机制。
方法：使用PCR阵列鉴定纤维性上皮病变和正常牙龈组织之间的差异表达基因（DEGs）。18个自噬相关（ATG）家族基因的表达水平，十二个B细胞淋巴瘤2（Bcl-2）家族基因，使用定量实时PCR（qRT-PCR）验证了11个半胱氨酸依赖性天冬氨酸定向蛋白酶（caspase）家族基因。通过免疫印迹分析测量微管相关蛋白轻链3(LC3)转化(LC3-I至LC3-II)来确定自噬诱导。
结果：PCR阵列鉴定了六个上调的基因，而没有基因表达水平显着降低。上调的基因是BCL2，BCL2L1，CXCR4，HSP90AA1，HSPA8和IGF1，它们都属于“自噬调节”组，而不是“自噬机制成分”组。qRT-PCR验证了BCL2、BCL2L1（也称为BCL-XL）的表达水平，BCL2L2(又称BCL-W)在纤维性腺中显著增高。没有观察到LC3-I到LC3-II的转化。
结论：本研究表明，Bcl-2和Bcl-xL协同介导牙龈细胞逃避凋亡，导致不受控制的扩散。此外，ATG家族基因未被激活,自噬不参与这个过程。
方法：使用PCR阵列鉴定纤维性上皮病变和正常牙龈组织之间的差异表达基因（DEGs）。18个自噬相关（ATG）家族基因的表达水平，十二个B细胞淋巴瘤2（Bcl-2）家族基因，使用定量实时PCR（qRT-PCR）验证了11个半胱氨酸依赖性天冬氨酸定向蛋白酶（caspase）家族基因。通过免疫印迹分析测量微管相关蛋白轻链3(LC3)转化(LC3-I至LC3-II)来确定自噬诱导。
结果：PCR阵列鉴定了六个上调的基因，而没有基因表达水平显着降低。上调的基因是BCL2，BCL2L1，CXCR4，HSP90AA1，HSPA8和IGF1，它们都属于“自噬调节”组，而不是“自噬机制成分”组。qRT-PCR验证了BCL2、BCL2L1（也称为BCL-XL）的表达水平，BCL2L2(又称BCL-W)在纤维性腺中显著增高。没有观察到LC3-I到LC3-II的转化。
结论：本研究表明，Bcl-2和Bcl-xL协同介导牙龈细胞逃避凋亡，导致不受控制的扩散。此外，ATG家族基因未被激活,自噬不参与这个过程。
