| Objectives:To investigate the effects of different concentrations of omentin-1 on surface markers of colon cancer stem cells in high glucose environment.And the effect of different concentrations of retinoid-1 on tumor suppressor mi RNA.Method:CD133+ colon cancer stem cells obtained by indirect immunomagnetic beads sorting in the early stage of the research group were taken as subjects,and CD133+ colon cancer stem cells were identified by immunocytochemistry.According to different research purposes,(1)Colon cancer stem cells were divided into 6 groups: Z0 group(without any intervention),Z1 group(1ug/m L omentin-1),Z2 group(2ug/m L omentin-1);G0 group(5.0g/m L glucose),G1 group(1ug/m L)omentin-1+5.0g/m L glucose),G2 group(2ug/m L omentin-1+5.0g/m L glucose).The m RNA and protein expressions of stem cell surface markers(CD133,CD44,ALDH1)in each group were detected by q PCR and Western blot After 24 hours of intervention.(2)Colon cancer stem cells were divided into three groups: Control group(without any treatment),Omentin 1 group(1ug/m L Omentin-1),Omentin 2 group(2ug/m L Omentin-1),The expression of tumor suppressor-related mi RNAs(mi R-126,mi R-34 a,mi R-143,mi R-145,mi R-342)was detected by q PCR after 24 hours of intervention.Results:1.Colon cancer stem cells were suspended growth in serum-free medium,then stem cell Spheres gradually formed and the spheres further increased and became round.CD133 Antigen was observed on the cell membrane surface of colon cancer stem cells by Immunocytochemical.2.Colon cancer stem cell surface marker expression changes(1)Expression of CD133 m RNA: Compared with Z0 group,the expression of CD133 m RNA in Z1 and Z2 groups was significantly decreased(P<0.05),and there was no significant difference between Z1 and Z2 groups(P>0.05).Compared with Z0 group,G0 The expression of CD133 m RNA was significantly increased in the group(P<0.05).Compared with the G0 group,the expression of CD133 m RNA in the G1 and G2 groups was significantly decreased(P<0.05).There was no significant difference in the expression of CD133 m RNA between the G1 and G2 groups(P>0.05).(2)The expression of CD44 m RNA: Compared with Z0 group and Z1 group,the expression of CD44 m RNA in Z2 group was significantly increased(P<0.05),but there was no significant difference in CD44 m RNA expression between Z0 group and Z1 group(P>0.05);Compared with G1 group,the expression of CD44 m RNA in G2 group was significantly increased(P<0.05),but there was no significant difference between G0 and G1 groups and between G2 and Z2 groups(P>0.05).(3)The expression of ALDH1 m RNA : There was no significant difference in ALDH1 m RNA expression between Z0,Z1,Z2 groups and G0,G1,G2 groups(P>0.05).(4)Expression of CD133 protein: Compared with Z0,the expression of CD133 protein in Z2 group was significantly decreased(P<0.05),but there was no significant difference in Z1 group(P>0.05);compared with G0 group,G133 and G2 group CD133 Protein expression was significantly decreased(P<0.05),but there was no significant difference between G1 and G2 groups(P>0.05).There was no significant difference in CD133 protein expression between G2 and Z2 groups(P>0.05).(5)The expression of CD44 and ALDH1 protein levels: There was no significant difference in protein expression between Z0,Z1,Z2 groups and G0,G1,G2 groups(P>0.05).3.Expression of tumor suppressor-related mi RNAs(mi R-126,mi R-34 a,mi R-143,mi R-145,mi R-342)Compared with the Omentin 1 group,the expression of mi RNA126,mi RNA 34 a,mi RNA 145 and mi RNA 342-5P in the Omentin 2 group was significantly increased(P<0.05),but there was no significant difference in the expression of the above mi RNA between the Control group and the Omentin1 group.(P>0.05);In addition,there was no significant difference in the expression of mi RNA 342-3P and mi RNA 143 between the Control group,the Omentin 1 group and the Omentin 2 group(P>0.05).Conclusion: 1.High glucose environment can up-regulate the expression of CD133 m RNA and protein in colon cancer stem cell surface markers 2.Omentin-1 can down-regulate the expression of CD133 m RNA and protein on colon cancer stem cell markers in high glucose environment,and up-regulate the expression of CD44 m RNA.3.Omentin-1 can promote the expression of tumor suppressor-associated mi RNA126,mi RNA 145,mi RNA 34 a and mi RNA 342-5P. |
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