[Objectives]To observe the effects of Cigu Xiaozhi Formula on miR-378a-3p expression and Hh signaling pathway in TGF-β1 induced and activated LX2 cells.[Methods]Cells were divided into control group,induction group,d...[Objectives]To observe the effects of Cigu Xiaozhi Formula on miR-378a-3p expression and Hh signaling pathway in TGF-β1 induced and activated LX2 cells.[Methods]Cells were divided into control group,induction group,drug-containing serum group,miR-378a-3p inhibitor group,and miR inhibitor NC group.CCK-8 method was used to detect the cell viability of each group,and flow cytometry was used to detect the apoptosis rate of each group.RT-qPCR was used to detect the expression of miR-378a-3p in each group s cells,and RT-qPCR and Western blot were used to detect mRNA and protein expression of Shh,Gli1,Gli2,Col-I,andα-SMA in each group s cells.[Results]Compared with the control group,the cell viability and expression of Shh,Gli1,Gli2,Col-I,andα-SMA mRNA and protein in induction group increased(P<0.01),while the expression of miR-378a-3p decreased(P<0.01).Compared with the induction group,the cell viability and expression of Shh,Gli1,Gli2,Col-I,α-SMA mRNA andα-SMA and Gli2 protein decreased in drug-containing serum group(P<0.05),while cell apoptosis rate and miR-378a-3p expression increased(P<0.01).In miR-378a-3p inhibitor group,cell viability and the expression of Shh,Gli1,Gli2,Col-I,α-SMA mRNA and Gli1,Gli2,α-SMA protein increased(P<0.05,P<0.01),while the apoptosis rate and miR-378a-3p expression decreased(P<0.05,P<0.01).[Conclusions]Cigu Xiaozhi Formula containing serum can upregulate miR-378a-3p expression and downregulate the expression of Gli2 andα-SMA in TGF-β1 induced LX2 cells,thereby inhibiting the activation of LX2 cells and exerting the effects of anti liver fibrosis.展开更多
Objective:To improve the prognosis of patients with gastric cancer(GC),more effective therapeutic targets are urgently needed.Increasing evidence indicates that miRNAs are involved in the progression of various tumors...Objective:To improve the prognosis of patients with gastric cancer(GC),more effective therapeutic targets are urgently needed.Increasing evidence indicates that miRNAs are involved in the progression of various tumors,and RAS-associated protein in the brain 31(RAB31)is upregulated and promotes the progression of multiple malignant tumors.Here,we focused on identifying RAB31-targeted miRNAs and elucidating their potential mechanism in the progression of GC.Methods:RAB31 and miR-378a-3p expression levels were detected in paired fresh GC tissues and GC cell lines.Bioinformatics analysis was used to predict the miRNAs targeting RAB31 and the relationships between RAB31 and other genes.Dual-luciferase reporter assays were applied to verify the targeted interaction relationship.CCK-8,colony formation,flow cytometry,wound healing,and Transwell assays were performed to assess the proliferation,apoptosis,migration,and invasion of GC cells.Tumorsphere formation assays were performed to assess the stemness of gastric cancer stem cells.Related proteins were detected by Western blot.Xenograft assays in nude mice were performed to explore the effect of miR-378a-3p in vivo.Results:We report the first evidence that miR-378a-3p is downregulated in GC,whereas its overexpression inhibits proliferation,invasion,and migration as well as promotes apoptosis in GC cells.Mechanistically,miR-378a-3p inhibits the progression of GC by directly targeting RAB31.Restoring RAB31 expression partially offsets the inhibitory effect of miR-378a-3p.Further research revealed that miR-378a-3p inhibits GLI1/2 in the Hedgehog signaling pathway and attenuates the stemness of gastric cancer stem cells.Finally,xenograft assays showed that miR-378a-3p inhibits GC tumorigenesis in vivo.Conclusions:MiR-378a-3p inhibits GC progression by directly targeting RAB31 and inhibiting the Hedgehog signaling pathway proteins GLI1/2.展开更多
基金Supported by Regional Fund Project of National Natural Science Foundation of China(81860821)Gansu Province Higher Education Innovation Ability Enhancement Project in 2019(2019B-104)Innovation and Entrepreneurship Fund for Graduate Students of Gansu University of Chinese Medicine(2022CX64).
文摘[Objectives]To observe the effects of Cigu Xiaozhi Formula on miR-378a-3p expression and Hh signaling pathway in TGF-β1 induced and activated LX2 cells.[Methods]Cells were divided into control group,induction group,drug-containing serum group,miR-378a-3p inhibitor group,and miR inhibitor NC group.CCK-8 method was used to detect the cell viability of each group,and flow cytometry was used to detect the apoptosis rate of each group.RT-qPCR was used to detect the expression of miR-378a-3p in each group s cells,and RT-qPCR and Western blot were used to detect mRNA and protein expression of Shh,Gli1,Gli2,Col-I,andα-SMA in each group s cells.[Results]Compared with the control group,the cell viability and expression of Shh,Gli1,Gli2,Col-I,andα-SMA mRNA and protein in induction group increased(P<0.01),while the expression of miR-378a-3p decreased(P<0.01).Compared with the induction group,the cell viability and expression of Shh,Gli1,Gli2,Col-I,α-SMA mRNA andα-SMA and Gli2 protein decreased in drug-containing serum group(P<0.05),while cell apoptosis rate and miR-378a-3p expression increased(P<0.01).In miR-378a-3p inhibitor group,cell viability and the expression of Shh,Gli1,Gli2,Col-I,α-SMA mRNA and Gli1,Gli2,α-SMA protein increased(P<0.05,P<0.01),while the apoptosis rate and miR-378a-3p expression decreased(P<0.05,P<0.01).[Conclusions]Cigu Xiaozhi Formula containing serum can upregulate miR-378a-3p expression and downregulate the expression of Gli2 andα-SMA in TGF-β1 induced LX2 cells,thereby inhibiting the activation of LX2 cells and exerting the effects of anti liver fibrosis.
文摘Objective:To improve the prognosis of patients with gastric cancer(GC),more effective therapeutic targets are urgently needed.Increasing evidence indicates that miRNAs are involved in the progression of various tumors,and RAS-associated protein in the brain 31(RAB31)is upregulated and promotes the progression of multiple malignant tumors.Here,we focused on identifying RAB31-targeted miRNAs and elucidating their potential mechanism in the progression of GC.Methods:RAB31 and miR-378a-3p expression levels were detected in paired fresh GC tissues and GC cell lines.Bioinformatics analysis was used to predict the miRNAs targeting RAB31 and the relationships between RAB31 and other genes.Dual-luciferase reporter assays were applied to verify the targeted interaction relationship.CCK-8,colony formation,flow cytometry,wound healing,and Transwell assays were performed to assess the proliferation,apoptosis,migration,and invasion of GC cells.Tumorsphere formation assays were performed to assess the stemness of gastric cancer stem cells.Related proteins were detected by Western blot.Xenograft assays in nude mice were performed to explore the effect of miR-378a-3p in vivo.Results:We report the first evidence that miR-378a-3p is downregulated in GC,whereas its overexpression inhibits proliferation,invasion,and migration as well as promotes apoptosis in GC cells.Mechanistically,miR-378a-3p inhibits the progression of GC by directly targeting RAB31.Restoring RAB31 expression partially offsets the inhibitory effect of miR-378a-3p.Further research revealed that miR-378a-3p inhibits GLI1/2 in the Hedgehog signaling pathway and attenuates the stemness of gastric cancer stem cells.Finally,xenograft assays showed that miR-378a-3p inhibits GC tumorigenesis in vivo.Conclusions:MiR-378a-3p inhibits GC progression by directly targeting RAB31 and inhibiting the Hedgehog signaling pathway proteins GLI1/2.
