研究目的
To identify and image miR-155 for early screening of lung cancer using targeted delivery of octreotide-conjugated chitosan-molecular beacon nanoparticles.
研究成果
CS-MB-OCT nanoparticles specifically bind to SSTR2 on tumor cells, enabling effective identification and imaging of miR-155 in lung cancer models. This provides a new method for early screening by dynamically monitoring disease progression through fluorescence intensity.
研究不足
The study may have limitations in generalizability to other cancer types or clinical translation, and potential optimizations in nanoparticle synthesis and targeting efficiency.
1:Experimental Design and Method Selection:
The study used molecular beacon technology and nanotechnology to synthesize CS-MB-OCT nanoparticles for targeted delivery and imaging. Models included lung cancer cells, subcutaneous xenografts in nude mice, and Lox-Stop-lox K-ras G12D transgenic mice to observe disease progression.
2:Sample Selection and Data Sources:
Human lung adenocarcinoma cells A549 and SPC-A1, prostate cancer cells PC-3, nude mice for xenografts, and transgenic mice for different disease stages. Cells were cultured in RPMI-1640 medium with 10% FBS.
3:List of Experimental Equipment and Materials:
Equipment includes confocal microscopy, Varioskan Flash, Malvern Zetasizer Nano, TEM, IVIS spectrum imaging system, flow cytometry. Materials include chitosan nanoparticles, molecular beacons, octreotide, glutaraldehyde, antibodies, dyes, and reagents from various suppliers.
4:Experimental Procedures and Operational Workflow:
Steps involved cell culture, animal model establishment, SSTR2 and miR-155 expression detection using immunofluorescence, qRT-PCR, synthesis and characterization of CS-MB-OCT, in vitro and in vivo fluorescence imaging, and statistical analysis.
5:Data Analysis Methods:
Statistical analysis used independent sample t-test and one-way ANOVA with p < 0.05 considered significant, data expressed as mean ± SD.
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Malvern Zetasizer Nano instrument
Malvern rn 3000HS
Malvern
Measurement of particle size
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RPMI-1640 medium
Hyclone
Cell culture medium
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fetal bovine serum
Invitrogen
Supplement for cell culture
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Cre adenovirus
Hanbio
Inducing lung adenocarcinoma in transgenic mice
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paraformaldehyde
Boster
Cell fixation
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SSTR2 primary antibodies
abcam
Detection of SSTR2 expression
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Alexa488-labeled goat anti-rabbit secondary antibody
ZSGB-bio
Immunofluorescence detection
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DAPI
Nuclear staining
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RNAiso plus reagent
TaKaRa
RNA extraction
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SYBR PCR Master Mix reagent kits
TaKaRa
qRT-PCR detection
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molecular beacon
Sangon Company
Detection of miR-155
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chitosan nanoparticle
Guanghan Hengyu Company
Vehicle for molecular beacon delivery
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octreotide
GL Biochem
Targeting ligand for SSTR2
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glutaraldehyde
Chemical cross-linker for conjugation
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OCT-FITC
Shanghai GL biochem company
Fluorescent labeling for imaging
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Varioskan Flash
Thermo
Detection of absorption spectra and fluorescence intensity
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TEM
Philips TECNAI 10
Philips
Observation of nanoparticle morphology
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Opti-MEM I medium
Life Technologies
Cell transfection medium
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Hoechst 33342
Beyotime
Nuclear staining
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flow cytometry
BD FACS Calibur
Becton
Analysis of transfection efficiency
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