Long non-coding RNA (ribonucleic acid) and application thereof in diagnosing/treating cervical cancer
A non-coding, cervical cancer technology, applied in the field of genetic engineering, can solve the problem that the function research of lncRNAs is not very thorough.
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Embodiment 1
[0073] Detection of the expression of RP11-7K24.3 in tissues and cells
[0074] Take 0.1g tissue, grind it sufficiently with liquid nitrogen (into powder) or 1-5×10 7 Discard the culture medium from the cells, and rinse twice with pre-cooled PBS. Add 1ml of Trizol lysate, pipette and mix with an enzyme-free pipette tip, let stand for 5min, and transfer the lysate into a pre-labeled 1.5ml enzyme-free centrifuge tube. Centrifuge at 7500g at 4°C for 5 minutes, take the supernatant and add 1 / 5 volume of chloroform, invert and mix for 30 seconds, and let stand for 2 minutes. Centrifuge at 12000g for 15min at 4°C. The solution is divided into three layers (aqueous phase-white precipitate-red organic matter), transfer the aqueous phase layer to a new 1.5ml centrifuge tube, try not to absorb the white precipitate. Add an equal volume of isopropanol, mix gently by inversion, and let stand for 5-10min. Centrifuge at 12000g for 10min at 4°C. Discard the supernatant, add 1ml of 75% e...
Embodiment 2
[0095] Effects of RP11-7K24.3 on the invasion and migration of cervical cancer cell SiHa
[0096] An important aspect in the pathogenesis of tumor cell invasion and metastasis in cervical cancer. SiHa was used as the research object, and lip2000 was used as the carrier to transfect RP11-7K24.3 interference sequence or RP11-7K24.3 plasmid to down-regulate or up-regulate the expression of RP11-7K24.3. Transwells experiments were used to study the effect of R P11-7K24.3 knockdown on the migration and invasion of SiHa cells. On the contrary, overexpression of RP11-7K24.3 had an effect on the migration and invasion abilities of SiHa cells. The results showed that the knockdown of RP11-7K24.3 promoted the migration ability of cervical cancer cell SiHa and inhibited the invasion ability compared with the control cells; the high expression of RP11-7K24.3 inhibited the cervical cancer cells compared with the control cells. Cell SiHa migration ability, inhibition of invasion ability (...
Embodiment 3
[0098] Effect of RP11-7K24.3 on Lymphangiogenesis of Cervical Cancer Cells
[0099] Lymphatic metastasis of tumor cells is an important aspect in the pathogenesis of cervical cancer. HLEC-P3 was used as the research object, and lip2000 was used as the carrier to transfect RP11-7K24.3 interference sequence or RP11-7K24.3 plasmid to down-regulate or up-regulate the expression of RP11-7K24.3. The effect of RP11-7K24.3 knockdown on the lymphangiogenesis of HLEC-P3 cells was studied by lymphangiogenesis assay. On the contrary, overexpression of RP11-7K24.3 has an effect on lymphangiogenesis of HLEC-P3 cells. The results showed that knockdown of RP11-7K24.3 promoted lymphangiogenesis of cervical cancer cell HLEC-P3 compared with control cells, and high expression of RP11-7K24.3 inhibited cervical cancer cell HLEC-P3 lymphangiogenesis compared with control cells. EC-P3 lymphangiogenesis capacity ( Figure 4 ). It can be seen that the low expression of RP11-7K24.3 promotes the lym...
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