Difference between revisions of "Part:BBa K3638003:Experience"

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=Applications of BBa_K3638003=
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__NOTOC__
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===effect of pEGFP-miR-155-sponge-1 in breast cancer cells===
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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MicroRNA-155 (miR-155) is a well-known oncogenic miRNA overexpressed in many human cancers, including breast cancer[5]. So we constructed pEGFP-miR-155-sponge-1 and try to test the possibility of detecting miR-155 expression by using this plasmid. In order to explore whether XIST correlates with pEGFP-miR-155 sensor in breast cancer cells with potential target sites. pEGFP–C1 (as negative controls), pEGFP-miR-155-sponge-1 (0.8 ug plasmids for each well) was transfected into human breast cells (MDA-MB 231 cells and MDA-MB 468 cells) in 24-well plate, respectively. After transfection, cells were examined under fluorescence microscopy (Fig.4 A, B, C, D and Table 1). The fluorescence of GFP was decreased in MDA-MB 231 cells and MDA-MB 468 cells transfected with pEGFP-miR-155-sponge-1 compared with controls (Fig. 5). In addition, we also measured the value of GFP fluorescence by plate reader (SpectraMax i3). The endogenous miR- 155 could inhibit the expression of GFP in cells transfected with pEGFP-miR-155-sponge-1 (Table 1 and Fig5). The results suggested the GFP value of cells transfected with pEGFP-miR-155-sponge-1 could show the different expression of miRNAs in cells.
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how you used this part and how it worked out.
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[[File:The value of GFP fluorescence in cells.png|600px]]
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<br/>Fig 4. The images of different breast cells transfected with different plasmids. (A). pEGFP–C1 was transfected in MDA-MB 468 cells. (B). pEGFP-miR-155-sponge-1 was transfected in MDA-MB 468 cells. (C). pEGFP–C1 was transfected in MDA-MB 231 cells. (D).pEGFP-miR-155-sponge-1 was transfected in MDA-MB 231 cells.
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===Applications of BBa_K3638003===
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[[File:The images of different breast cells transfected with different plasmids.png|500px]]
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[[File:Fig5. The value of GFP fluorescence in cells. Different cells were transfected with pEGFP-C1 or pEGFP-miR-155-sponge-1 for 48 h.png|600px]]
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<br/>Fig5. The value of GFP fluorescence in cells. Different cells were transfected with pEGFP-C1 or pEGFP-miR-155-sponge-1 for 48 h.
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padding: 2px;">(图片来源: NASA's Imagine the Universe)</div>
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===User Reviews===
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<partinfo>BBa_K3638003 AddReview number</partinfo>
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<I>Username</I>
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Enter the review inofrmation here.
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Latest revision as of 15:13, 3 October 2020

Applications of BBa_K3638003

effect of pEGFP-miR-155-sponge-1 in breast cancer cells

MicroRNA-155 (miR-155) is a well-known oncogenic miRNA overexpressed in many human cancers, including breast cancer[5]. So we constructed pEGFP-miR-155-sponge-1 and try to test the possibility of detecting miR-155 expression by using this plasmid. In order to explore whether XIST correlates with pEGFP-miR-155 sensor in breast cancer cells with potential target sites. pEGFP–C1 (as negative controls), pEGFP-miR-155-sponge-1 (0.8 ug plasmids for each well) was transfected into human breast cells (MDA-MB 231 cells and MDA-MB 468 cells) in 24-well plate, respectively. After transfection, cells were examined under fluorescence microscopy (Fig.4 A, B, C, D and Table 1). The fluorescence of GFP was decreased in MDA-MB 231 cells and MDA-MB 468 cells transfected with pEGFP-miR-155-sponge-1 compared with controls (Fig. 5). In addition, we also measured the value of GFP fluorescence by plate reader (SpectraMax i3). The endogenous miR- 155 could inhibit the expression of GFP in cells transfected with pEGFP-miR-155-sponge-1 (Table 1 and Fig5). The results suggested the GFP value of cells transfected with pEGFP-miR-155-sponge-1 could show the different expression of miRNAs in cells. The value of GFP fluorescence in cells.png
Fig 4. The images of different breast cells transfected with different plasmids. (A). pEGFP–C1 was transfected in MDA-MB 468 cells. (B). pEGFP-miR-155-sponge-1 was transfected in MDA-MB 468 cells. (C). pEGFP–C1 was transfected in MDA-MB 231 cells. (D).pEGFP-miR-155-sponge-1 was transfected in MDA-MB 231 cells. The images of different breast cells transfected with different plasmids.png Fig5. The value of GFP fluorescence in cells. Different cells were transfected with pEGFP-C1 or pEGFP-miR-155-sponge-1 for 48 h.png
Fig5. The value of GFP fluorescence in cells. Different cells were transfected with pEGFP-C1 or pEGFP-miR-155-sponge-1 for 48 h.