研究目的
To investigate the effect and mechanism of 630-nm laser on human lung adenocarcinoma cell xenograft model in nude mice mediated by hematoporphyrin derivatives (HPD) and provide theoretical basis for clinical photodynamic therapy (PDT).
研究成果
The 630-nm laser mediated by hematoporphyrin derivatives can significantly inhibit the growth of human lung adenocarcinoma xenograft tumor in nude mice, the mechanism of which is related to the inhibition of tumor angiogenesis by down-regulating VEGF and HIF-1α gene expression, and the promotion of tumor apoptosis by up-regulating Bax, Caspase-3, and down-regulating Bcl-2 gene expression.
研究不足
The exact signal transduction pathway involved in the mechanism of action of the 630-nm laser mediated by HPD on the transplanted tumor in nude mice needs to be further studied.
1：Experimental Design and Method Selection:
The study involved establishing a human lung adenocarcinoma cell xenograft model in nude mice, dividing them into four groups (control, pure photosensitizer, pure irradiation, and photodynamic treatment groups), and comparing tumor volume growth and inhibition rates. HE and TUNEL staining were used for pathological observation and apoptosis detection, respectively. RT-PCR and Western blot were employed to detect the expression levels of angiogenesis and apoptosis-related factors.
2：Sample Selection and Data Sources:
Twenty-four female BALB/c nude mice were used to establish the xenograft model with human lung adenocarcinoma A549 cells.
3：List of Experimental Equipment and Materials:
RPMI-1640 medium, fetal bovine serum, trypsin, double antibody, dimethyl sulfoxide (DMSO), HE staining kit, TUNEL cell apoptosis detection kit, RIPA lysis buffer, BCA Protein Assay Kit, reverse transcription kit, SYBR Green quantitative PCR kit, Western blotting primary and secondary antibody diluent, PVDF membrane, ECL chemiluminescence kit, monoclonal antibodies for Caspase-3, Bax, Bcl-2, VEGF, HIF-1α, and GAPDH, LED-IB photodynamic therapy instrument.
4：Experimental Procedures and Operational Workflow:
The xenograft model was established, and the mice were treated according to their group assignments. Tumor volume was measured, and samples were collected for HE and TUNEL staining, RT-PCR, and Western blot analysis.
5：Data Analysis Methods:
Statistical analysis was performed using SPSS23.0 software, with data expressed as mean ± standard deviation. Comparisons were made using single factor analysis of variance and LSD-t test.
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RPMI-1640 medium
Hyclone Company
Culture medium for cell growth
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fetal bovine serum
Hyclone Company
Supplement for cell culture medium
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trypsin
Hyclone Company
Enzyme for cell detachment
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double antibody
100 U/ml penicillin and 0.1 mg/ml streptomycin
Hyclone Company
Antibiotic for cell culture
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dimethyl sulfoxide
Sigma Company
Solvent
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HE staining kit
Roche Company
Pathological observation of tumor sections
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TUNEL cell apoptosis detection kit
Roche Company
Detection of apoptosis in tumor tissues
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RIPA lysis buffer
Beijing Solarbio Company
Protein extraction
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BCA Protein Assay Kit
Beijing Solarbio Company
Protein concentration determination
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reverse transcription kit
TaKaRa Company
cDNA synthesis
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SYBR Green quantitative PCR kit
TaKaRa Company
Quantitative PCR
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Western blotting primary and secondary antibody diluent
Wuhan BOSTER biological company
Antibody dilution for Western blot
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PVDF membrane
MILLIPORE Company
Protein transfer in Western blot
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ECL chemiluminescence kit
MILLIPORE Company
Detection of proteins in Western blot
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LED-IB photodynamic therapy instrument
Red light 630 nm; Output power: 80 mW/cm2
Wuhan Yage Photoelectric Technology Co. LTD
Photodynamic therapy
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