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ShRNA (short hairpin ribonucleic acid) for knocking down PXYLP1 gene expression, lentiviral vector as well as construction method and application of lentiviral vector

A lentiviral vector and gene expression technology, applied in the field of short hairpin RNA-mediated knockdown of tongue squamous cell carcinoma PXYLP1, can solve the problems of poor selectivity, low targeting, and large side effects of chemotherapy drugs, and achieve auxiliary diagnosis and promote Apoptotic effect

Active Publication Date: 2022-05-10
XUZHOU CENT HOSPITAL
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Problems solved by technology

[0003] At present, compounds or natural extracts are widely used in the treatment of squamous cell carcinoma of the tongue, such as CN110680815 A and CN 111773218 A, but the application of small interfering RNA biological drugs in the preparation of squamous cell carcinoma of the tongue is less, and the side effects of chemotherapy drugs Large, poor selectivity, low targeting, limited application

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  • ShRNA (short hairpin ribonucleic acid) for knocking down PXYLP1 gene expression, lentiviral vector as well as construction method and application of lentiviral vector
  • ShRNA (short hairpin ribonucleic acid) for knocking down PXYLP1 gene expression, lentiviral vector as well as construction method and application of lentiviral vector
  • ShRNA (short hairpin ribonucleic acid) for knocking down PXYLP1 gene expression, lentiviral vector as well as construction method and application of lentiviral vector

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Embodiment Construction

[0043] The inventive method of the present invention is described and illustrated in detail below in conjunction with specific examples. Its content is an explanation of the present invention rather than limiting the protection scope of the present invention.

[0044]Our research found that designing the interference target sequence for the PXYLP1 gene, packaging and constructing lentivirus, and knocking down the PXYLP1 gene will significantly affect the proliferation of tongue squamous cell carcinoma cells CAL-27 cells and SCC-25 cells, and affect the growth and formation of cells. ability, and promote cell apoptosis, and was verified by in vivo experiments. It shows that the gene may regulate the occurrence and development of tumors, and may become a potential therapeutic target for tongue squamous cell carcinoma.

[0045] (1) Experimental method

[0046] 1. Gene information

[0047] The online resource TCGA database (http: / / ualcan.path.uab.edu / ) was used to generate diff...

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Abstract

The invention discloses shRNA (short hairpin ribonucleic acid) for knocking down PXYLP1 gene expression, a lentiviral vector and a construction method and application thereof. RNA interference target design and double-stranded DNA oligo preparation are performed, the double-stranded DNA oligo is connected with a linearized vector, a connection product is converted into an escherichia coli competent cell for positive cloning, a short hairpin RNA lentiviral vector is constructed, expression of a TSCC endogenous PXYLP1 gene is knocked down, and the PXYLP1 gene expression is knocked down. The proliferation capacity of the TSCC cells is inhibited, and the apoptosis of the TSCC cells is induced, so that in-vivo tumor growth is inhibited.

Description

technical field [0001] The invention relates to the field of cancer treatment, in particular to a short hairpin RNA-mediated knockdown method of PXYLP1 in tongue squamous cell carcinoma cells and its application. Background technique [0002] Tongue squamous cell carcinoma (TSCC) is one of the most common malignant tumors with high mortality and morbidity. Its development is a multistep process involving genetic alterations and epigenetic modifications. TSCC is a causative factor of many variables, accumulation of multiple genetic alterations and environmental factors, such as tobacco use, alcohol consumption, chronic inflammation, and human papillomavirus (HPV) infection, among others, and some recent reports suggest that TSCC in young adults is increasing in incidence , especially in women, the rate of distant metastasis is higher and the prognosis is poorer. Therefore, TSCC caused by different risk factors may have different molecular basis, involving the loss of cell c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/867A61K31/713A61P1/02A61P25/00
CPCC12N15/1137C12N15/86A61K31/713A61P1/02A61P25/00C12N2310/14C12N2740/15043Y02A50/30
Inventor 孟箭周霖李欣然顾徐嘉陈霖
Owner XUZHOU CENT HOSPITAL
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