目的研究miRNA-195在宫颈癌组织和细胞中的表达,分析其对宫颈癌细胞增殖、迁移的影响和作用机制。方法检测35例临床宫颈癌组织及其对应癌旁正常组织中miRNA-195,通过Kaplan-Meier Plotter数据库分析其与宫颈癌患者预后的关系;采用细胞...目的研究miRNA-195在宫颈癌组织和细胞中的表达,分析其对宫颈癌细胞增殖、迁移的影响和作用机制。方法检测35例临床宫颈癌组织及其对应癌旁正常组织中miRNA-195,通过Kaplan-Meier Plotter数据库分析其与宫颈癌患者预后的关系;采用细胞增殖实验和划痕迁移实验验证miRNA-195对宫颈癌siHa,Hela细胞增殖、迁移的影响;通过microRNA数据库预测miRNA-195的靶基因,双荧光素酶基因实验验证靶向结合关系;qRT-PCR验证miRNA-195对靶基因的调控;分析宫颈癌中靶基因的表达作用及与miRNA-195的相关性,通过细胞回补实验验证miRNA-195是否通过靶向调控蛋白表达在宫颈癌中发挥功能。结果宫颈癌组织中miRNA-195相对表达低于癌旁正常组织(21.03±5.17 vs 40.67±7.92),差异有统计学意义(t=12.285,P<0.001),且具有低表达预后差的临床特征(Logrank P=0.032)。过表达miRNA-195抑制了宫颈癌细胞的增殖(t=6.725~21.433,均P<0.01)和迁移速率(t=12.443,16.749,均P<0.001)。CCND2和MYB是miRNA-195的靶基因,过表达miRNA-195显著抑制了CCND2和MYB mRNA的蛋白表达(P<0.01)。宫颈癌组织中CCND2较癌旁正常组织显著高表达(52.67±4.79 vs 39.86±6.39),差异有统计学意义(t=12.453,P<0.001);MYB较癌旁正常组织显著高表达(43.06±6.43 vs 22.07±6.85),差异有统计学意义(t=13.217,P<0.001);且分别与miRNA-195表达呈负相关(r=-0.726,-0.592,均P<0.05)。过表达CCND2和MYB显著促进了宫颈癌细胞的增殖和迁移速率,敲低CCND2和MYB表达则得到与之相反的结果(F=144.947,875.160,均P<0.001);在过表达miRNA-195细胞中分别回补过表达CCND2和MYB后细胞增殖、迁移速率基本回归到正常水平。结论miRNA-195可通过靶向调控CCND2和MYB表达抑制宫颈癌癌细胞的增殖和迁移,进而参与宫颈癌的发生发展。展开更多
Objective:To explore the effect of propofol(Prof)on the proliferation,migration and invasion of human gastric cancer cell MGC-803 and its molecular mechanism.Methods:The MTT method was used to study the effects of Pro...Objective:To explore the effect of propofol(Prof)on the proliferation,migration and invasion of human gastric cancer cell MGC-803 and its molecular mechanism.Methods:The MTT method was used to study the effects of Prof with different doses and durations on the viability of MGC-803 cells.Hoechst 33258 staining and electron microscopy were used to detect the effects of Prof on MGC-803 cell apoptosis.Transwell experiments were used to detect the effects of Prof on the migration and invasion of MGC-803 cells.RT-PCR detects the effect of Prof on the expression of miR-195 in MGC-803 cells,and Western Blot detects the effect of Prof on the protein expression of JAK/STAT signaling pathway.Results:Compared with 0μg/ml Prof,5μg/ml,10μg/ml and 20μg/ml Prof treatment with 24h,48h and 72h can significantly reduce cell viability(P<0.05).Compared with the Control group,the percentage of Hoechst 33258 staining positive cells in the Prof group and the apoptosis rate under the electron microscope were significantly increased(P<0.05).Compared with the Control group,the cell migration rate and invasion rate of the Prof group were significantly reduced(P<0.05).Compared with the Control group,the expression of miRNA-195 in the Prof group cells was increased significantly(P<0.05).Compared with the Control group,the activity of p-Jak1 and p-STAT3 proteins in the Prof group were significantly reduced(P<0.05).Conclusion:Prof can reduce the cell viability,migration and invasion of gastric cancer cell MGC-803,and promote its apoptosis.Its mechanism may be related to the promotion of miR-195 expression and inhibition of JAK/STAT signal pathway activity.展开更多
文摘目的研究miRNA-195在宫颈癌组织和细胞中的表达,分析其对宫颈癌细胞增殖、迁移的影响和作用机制。方法检测35例临床宫颈癌组织及其对应癌旁正常组织中miRNA-195,通过Kaplan-Meier Plotter数据库分析其与宫颈癌患者预后的关系;采用细胞增殖实验和划痕迁移实验验证miRNA-195对宫颈癌siHa,Hela细胞增殖、迁移的影响;通过microRNA数据库预测miRNA-195的靶基因,双荧光素酶基因实验验证靶向结合关系;qRT-PCR验证miRNA-195对靶基因的调控;分析宫颈癌中靶基因的表达作用及与miRNA-195的相关性,通过细胞回补实验验证miRNA-195是否通过靶向调控蛋白表达在宫颈癌中发挥功能。结果宫颈癌组织中miRNA-195相对表达低于癌旁正常组织(21.03±5.17 vs 40.67±7.92),差异有统计学意义(t=12.285,P<0.001),且具有低表达预后差的临床特征(Logrank P=0.032)。过表达miRNA-195抑制了宫颈癌细胞的增殖(t=6.725~21.433,均P<0.01)和迁移速率(t=12.443,16.749,均P<0.001)。CCND2和MYB是miRNA-195的靶基因,过表达miRNA-195显著抑制了CCND2和MYB mRNA的蛋白表达(P<0.01)。宫颈癌组织中CCND2较癌旁正常组织显著高表达(52.67±4.79 vs 39.86±6.39),差异有统计学意义(t=12.453,P<0.001);MYB较癌旁正常组织显著高表达(43.06±6.43 vs 22.07±6.85),差异有统计学意义(t=13.217,P<0.001);且分别与miRNA-195表达呈负相关(r=-0.726,-0.592,均P<0.05)。过表达CCND2和MYB显著促进了宫颈癌细胞的增殖和迁移速率,敲低CCND2和MYB表达则得到与之相反的结果(F=144.947,875.160,均P<0.001);在过表达miRNA-195细胞中分别回补过表达CCND2和MYB后细胞增殖、迁移速率基本回归到正常水平。结论miRNA-195可通过靶向调控CCND2和MYB表达抑制宫颈癌癌细胞的增殖和迁移,进而参与宫颈癌的发生发展。
基金Science and Technology Planning of Xi'an[Project No.:20YXYJ0002(9)].
文摘Objective:To explore the effect of propofol(Prof)on the proliferation,migration and invasion of human gastric cancer cell MGC-803 and its molecular mechanism.Methods:The MTT method was used to study the effects of Prof with different doses and durations on the viability of MGC-803 cells.Hoechst 33258 staining and electron microscopy were used to detect the effects of Prof on MGC-803 cell apoptosis.Transwell experiments were used to detect the effects of Prof on the migration and invasion of MGC-803 cells.RT-PCR detects the effect of Prof on the expression of miR-195 in MGC-803 cells,and Western Blot detects the effect of Prof on the protein expression of JAK/STAT signaling pathway.Results:Compared with 0μg/ml Prof,5μg/ml,10μg/ml and 20μg/ml Prof treatment with 24h,48h and 72h can significantly reduce cell viability(P<0.05).Compared with the Control group,the percentage of Hoechst 33258 staining positive cells in the Prof group and the apoptosis rate under the electron microscope were significantly increased(P<0.05).Compared with the Control group,the cell migration rate and invasion rate of the Prof group were significantly reduced(P<0.05).Compared with the Control group,the expression of miRNA-195 in the Prof group cells was increased significantly(P<0.05).Compared with the Control group,the activity of p-Jak1 and p-STAT3 proteins in the Prof group were significantly reduced(P<0.05).Conclusion:Prof can reduce the cell viability,migration and invasion of gastric cancer cell MGC-803,and promote its apoptosis.Its mechanism may be related to the promotion of miR-195 expression and inhibition of JAK/STAT signal pathway activity.
