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A spio-shRNA molecular probe with dual functions of mr gene imaging and targeted therapy

A molecular probe and targeted therapy technology, applied in the field of medicine and medicine, can solve the problems of poor effect and large side effects, and achieve the effect of effective display and intervention and induction of cell apoptosis.

Inactive Publication Date: 2016-01-13
THE FIRST AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the imaging diagnostic methods currently used in clinical practice are all aimed at morphological imaging after the tumor has formed a substantial mass. At this time, the patient is already in the middle and late stages of the disease course. Common treatment modes such as surgery, chemotherapy, and radiotherapy have large side effects. not effectively

Method used

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  • A spio-shRNA molecular probe with dual functions of mr gene imaging and targeted therapy
  • A spio-shRNA molecular probe with dual functions of mr gene imaging and targeted therapy
  • A spio-shRNA molecular probe with dual functions of mr gene imaging and targeted therapy

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Preparation of SPIO-shRNA Molecular Probes

[0032] ① Preparation of SPIO: It is completed by co-precipitation method, and the reaction principle is: Fe 2+ +2Fe 3+ +8OH - → Fe 3 o 4 +4H 2 O.

[0033] Precisely weigh FeCl 3 ·6H 2 O1.17g with FeCl 2 4H 2 O0.43g (Fe 3+ and Fe 2+Molar ratio = 1:1.75) and dextran T-32.5g were dissolved in 40ml ultrapure water, and ammonia water was slowly added dropwise in the solution to adjust the pH value, so that the iron salt was combined with OH-, and then placed in a water bath at 80°C Stirring at 450r / min for 60min, during which nitrogen gas was kept flowing, after the reaction was completed, the pH was adjusted to 10 with acetic acid, and the centrifuge was centrifuged at 12000r. The bag was dialyzed for 24 hours, filtered through a microporous membrane, and then sonicated for 5 minutes to obtain dextran-modified SPIO. The sample was measured by transmission electron microscope and atomic force microscope: the core part...

Embodiment 2

[0037] Embodiment 2 performance index measurement

[0038] Performance:

[0039] (1) Biological and physical property testing:

[0040] 1. Detection of the binding rate of the SPIO-PLL complex to the plasmid pGenesil1-shRNA: As the concentration of SPIO-PLL increases, the binding rate to the plasmid gradually increases. When the ratio of the two reaches 6:1, it enters a plateau (see figure 1 ), suggesting that the combination of the two is close to saturation, therefore, the best mass ratio is 6-8:1.

[0041] 2. Gel retardation experiment: As shown in the figure below, under neutral conditions, the control group (lane 1 is the plasmid pGenesil1-shRNA, and lane 3 is the SPIO-PLL complex) can see white bright bands swimming out, In the experimental group (molecular probe in lane 2), there is only light in the spotting well, and no plasmid band is seen (see figure 2 ).

[0042] 3. Zeta potential detection: when PH=7, the zeta potential of SPIO is (-14.8±0.35) mV, the zeta po...

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Abstract

The invention relates to an SPIO-shRNA (superparamagnetic iron oxide-short hair ribonucleic acid) molecular probe with dual actions of MR (magnetic resonance) gene imaging and targeting therapy. The SPIO-shRNA comprises an SPIO nanoparticle and a pGenesil1-shRNA plasmid, wherein the sequence of the plasmid is AGTGTGTAACGGAATAGGT. The molecular probe can be used for MR (magnetic resonance) scanning gene imaging and detecting the specificity and targeting property on one hand, and for observing that the probe inhibits the EGFR (epidermal growth factor receptor) expression in cells, inducing cell apoptosis, regulating and controlling cell cycle redistribution and the like on the other hand, thus realizing targeting gene therapy.

Description

technical field [0001] The invention belongs to the field of medicine and medicine, and in particular relates to a SPIO-shRNA molecular probe with dual functions of MR gene imaging and tumor targeting therapy. Background technique [0002] Malignant tumors are common and frequently-occurring diseases that threaten the health of urban and rural people in my country, and are increasing year by year. Early, specific diagnosis and targeted treatment of malignant tumors are the key to improving survival rates and quality of life. However, the imaging diagnostic methods currently used in clinical practice are all aimed at morphological imaging after the tumor has formed a substantial mass. At this time, the patient is already in the middle and late stages of the disease course. Common treatment modes such as surgery, chemotherapy, and radiotherapy have large side effects. not effectively. [0003] RNA interference (RNAi) is a common natural phenomenon in eukaryotes. Compared with...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K49/12A61K48/00A61P35/00
Inventor 文明刘雨村文曦琳李易葛晓东邓小林李少林
Owner THE FIRST AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIVERSITY
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