研究目的
To evaluate the antitumor effect of sinoporphyrin sodium mediated photodynamic therapy (DVDMS-PDT) against human colorectal cancer (CRC) and to investigate the role of autophagy in its effect.
研究成果
DVDMS-PDT is more effective than PF-PDT in inhibiting HCT116 cells, inducing both apoptosis and autophagy. Autophagy inhibition with chloroquine enhances antitumor efficiency, suggesting a protective role of autophagy. DVDMS is a promising photosensitizer, and combined therapy could be feasible for CRC treatment.
研究不足
The study is limited to HCT116 colorectal cancer cells and may not generalize to other cancer types. The in vivo model uses nude mice, which might not fully replicate human physiology. Autophagy mechanisms and long-term effects require further investigation.
1：Experimental Design and Method Selection:
The study used in vitro and in vivo models to assess DVDMS-PDT effects, including cell viability, apoptosis, autophagy, and tumor growth inhibition. Methods included flow cytometry, western blotting, TEM, and animal xenograft models.
2：Sample Selection and Data Sources:
HCT116 human colorectal cancer cells were used, obtained from Cell Resource Center, Chinese Academy of Medical Science. Animal models used BALB/C nude mice.
3：List of Experimental Equipment and Materials:
Equipment included flow cytometer (FACSCalibur, BD, USA), transmission electron microscope (H-7650, HITACHI, Japan), laser device (630 nm, Xingda photoelectric medical instrument Co., Ltd.), and various chemicals like DVDMS, chloroquine, antibodies.
4：Experimental Procedures and Operational Workflow:
Cells were cultured, treated with DVDMS or Photofrin, exposed to laser light, and analyzed for viability, apoptosis, autophagy markers, and protein expression. In vivo, mice were injected with HCT116 cells, treated with PS and light, and tumors were measured and analyzed.
5：Data Analysis Methods:
Statistical analysis used one-way ANOVA with Turkey's test, performed with GraphPad Prism 7.0 software.
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transmission electron microscope
H-7650
HITACHI
Used to observe autophagic vacuole structures in cells.
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flow cytometer
FACSCalibur
BD
Used to detect intracellular DVDMS uptake and quantify apoptosis rates in cells.
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laser device
630 nm semiconductor laser
Xingda photoelectric medical instrument Co., Ltd.
Used as a light source for photodynamic therapy activation.
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CCK-8 assay kit
Dojindo Laboratories
Used to measure cell viability.
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FITC Annexin V and Propidium Iodide Kit
Biolegend
Used to quantify cell apoptosis by flow cytometry.
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MDC stain
Leagene Biotechnology
Used to stain and quantify autophagic vacuoles in cells.
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RIPA lysis buffer
Biosharp
Used to isolate total proteins from cells for western blotting.
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BCA detection kit
Beyotime
Used to determine protein concentrations.
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PVDF membranes
Millipore
Used in western blotting for protein transfer.
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ECL detection system
Tanon
Used to visualize proteins in western blotting.
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TUNEL in situ cell death detection kit
Roche
Used to identify apoptotic cells in tissue sections.
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