phytochlorin and Liver-Neoplasms

MicroRNAs (miRNAs) is the tiny and highly conserved noncoding RNAs, regulate gene expression at the post-transcriptional level by binding to the 3'-UTR of target mRNAs. Several studies found that miR-195 plays an unavoidable role in the regulation of cell proliferation, cycle and apoptosis in hepatocellular carcinoma (HCC). Here, we constructed miR-195 and Chlorine e6 (Ce6) co-loading NBs (nanobubbles), making use of NBs as carriers to deliver miR-195 and Ce6 to mouse tumor models. Our results showed that the binding between PD-1 and PD-L1 was blocked by upregulating miR-195 expression. The analysis of CTL (Cytotoxic T Cell) immune activity in the treatment group was higher than the control group. Simultaneously, Ce6 was used as sonosensitizer to induce SDT (sonodynamic therapy) and trigger ICD (immunogenic cell death) of tumor cell via generation of ROS. Recent studies have found that ICD may further enhance anti-tumor immunity against PD-L1. Results indicated that combination treatment effectively stimulated infiltration of T cell and the activation of natural killer (NK) cells as well as the maturation of dendritic cells (DCs), and the combination treatment group exibited the highest CTL killing activity. These results indicate that a stronger antitumor immunity was triggered via combination of SDT-induced tumor cell ICD and immune checkpoint blockade of PD-1/PD-L1 mediated by upregulation of miR-195. In conclusion, we have successfully constructed an efficient delivery system with great potential to provide a new strategy for synergistic immunotherapy.

Topics: Animals; B7-H1 Antigen; Carcinoma, Hepatocellular; Immune Checkpoint Inhibitors; Immunity; Liver Neoplasms; Mice; MicroRNAs; Nanoparticle Drug Delivery System; Programmed Cell Death 1 Receptor

A promising direction in Biopharmaceuticals is the development of specific peptide-based systems to improve drug delivery. This approach may increase tumor specificity and drug penetration into the target cell. Similar systems have been designed for several antitumor drugs. However, for photodynamic therapy drugs, such studies are not yet enough. Previously, we have developed a method of inclusion of chlorin e6 (Ce6), a photosensitizer used in photodynamic therapy, in phospholipid nanoparticles with a diameter of up to 30 nm, and reported an increase in its effectiveness in the experiments in vivo. In this work, we propose to modify a previously developed delivery system for Ce6 by the addition of cell-penetrating (R7) and/or targeting NGR peptides. The interaction of the compositions developed with HepG2 and MCF-7 tumor cells is shown. The expression of CD13 protein with affinity to NGR on the surface of these cells has been studied using flow cytometry. The expression of this protein on the HepG2 cells and its absence on MCF-7 was demonstrated. After incubation of tumor cells with the resulting Ce6 compositions, we evaluated the cellular accumulation, photoinduced, and dark cytotoxicity of the drugs. After irradiation, the highest level of cytotoxicity was observed when R7 peptide was added to the system, either alone or in combination with NGR. In addition to R7, the NGR-motif peptide increased the internalization of Ce6 in HepG2 cells without affecting its photodynamic activity. In this work we also discuss possible mechanisms of action of the cell-penetrating peptide when attached to phospholipid nanoparticles.

Topics: Breast Neoplasms; Carcinoma, Hepatocellular; CD13 Antigens; Cell Survival; Cell-Penetrating Peptides; Chlorophyllides; Drug Carriers; Drug Compounding; Hep G2 Cells; Humans; Liver Neoplasms; MCF-7 Cells; Nanoparticles; Oligopeptides; Phospholipids; Photochemotherapy; Photosensitizing Agents; Porphyrins

Immune checkpoint inhibitors (ICIs) and sonodynamic therapy (SDT) are types of immunotherapy. In order to combine soluble programmed cell death protein 1 (sPD-1)-mediated immune checkpoint therapy and chlorin e6 (Ce6)-assisted SDT, nanobubbles (NBs) were generated to simultaneously load sPD-1 and Ce6.. The sPD-1/Ce6-NBs, which were prepared by thin-film hydration and mechanical oscillation, had a stable physical condition, and delivered sPD-1 and Ce6 in a targeted manner. NBs could strengthen tumor suppression by increasing tumor-targeting accumulation of Ce6 and sPD-1, and by inducing ultrasound-targeted NB destruction. A mouse H22 cell hepatoma xenograft model was used to evaluate the synergetic immunotherapeutic effect and mechanism of sPD-1/Ce6-NBs.. By observing the tumor inhibition rate, tissue and cell apoptosis, apoptosis-related genes and protein expression, the best immunotherapeutic effect was exhibited by the sPD-1/Ce6-NBs group. The immunotherapeutic mechanism initially demonstrated that when tumor cells were transfected by sPD-1 delivered by NBs, which downregulated the expression of programmed death-ligand 1 (PD-L1) in tumor cells, and blocked the PD-1/PD-L1 signaling pathway, which improved T-cell-mediated tumor inhibition. Furthermore, ICIs combined with SDT induced immunogenic cell death by translocating calreticulin to the cell surface and then synergistically enhancing antitumor immune responses.. In conclusion, sPD-1/Ce6-NBs were successfully designed. Ultrasound-mediated sPD-1/Ce6-NBs are potentially effective delivery systems for combination immunotherapy of hepatocellular carcinoma.

Topics: Animals; Apoptosis; Carcinoma, Hepatocellular; Chlorophyllides; Humans; Immunotherapy; Liver Neoplasms; Male; Mice, Inbred BALB C; Nanoparticles; Porphyrins; Programmed Cell Death 1 Receptor; RNA, Messenger; T-Lymphocytes; Tumor Burden

Hepatocellular carcinoma (HCC) is easy to relapse after resection for its lack of anti-tumor immunity due to pro-tumorigenesis by promoting M2 type macrophage polarization. Recent studies have shown that exosomes are closely related to the occurrence and development of HCC. Antigenic exosomes from HCC are able to polarize into alternatively activated macrophages M2, but do not stimulate M1 macrophages polarization. Iron oxide nanoparticles (IONs) have been demonstrated to be able to promote M1 macrophages polarization. This research was to explore exosomes as vehicles to synergize with pegylated IONs loaded with chlorin e6 (PIONs@E6) to enhance their immunity against HCC via promoting M1 macrophages polarization.. PIONs@E6 was synthesized and then characterized by chemico-physical analysis, transmission electron microscope (TEM), respectively. After characterization of PIONs-contained exosomes by TEM, and then the exosomal surface specific molecules CD9 and CD63 were determined by Western Blotting assay. Markers of M1 macrophage polarization in vitro and in vivo were analyzed by enzyme linked immunosorbent assay (ELISA) and flow cytometry, respectively. Intracellular reactive oxygen species (ROS) in macrophages were analyzed using a Spectra Max fluorescence microplate reader. Inhibitory effect of PIONs-contained exosomes on HCC was evaluated by monitoring tumor growth in an in vivo xenograft mice model.. PIONs@E6 showed good water solubility with a core diameter around 10 nm and a hydrate diameter around 37 nm. The expression of exosome specific markers CD9 and CD63 was kept at a high level. PIONs-contained exosomes can dose-dependently promote M1 macrophages polarization in vitro and in vivo. Of note, PIONs-contained exosomes could initiate a significantly higher level of ROS in macrophages and remarkably inhibit the tumor growth in mice bearing HCC xenograft.. Exosomes as vehicles could be synergized with PIONs@E6 to enhance their immunity against HCC via promoting M1 macrophages polarization.

Topics: Animals; Biomarkers, Tumor; Carcinoma, Hepatocellular; Chlorophyllides; Exosomes; Female; Humans; Liver Neoplasms; Macrophage Activation; Macrophages; Magnetic Iron Oxide Nanoparticles; Mice, Inbred ICR; Reactive Oxygen Species; Tetraspanin 29; Tetraspanin 30

It is important to explore effective treatment for liver cancer. Photodynamic therapy (PDT) is a novel technique to treat liver cancer, but its clinical application is obstructed by limited depth of visible light penetration into tissue. The near-infrared (NIR) photosensitizer is a potential solution to the limitations of PDT for deep tumor tissue treatment.. We aimed to investigate 808 nm NIR light-excited UCNPs@mSiO. In our study, 808 nm NIR light-excited upconversion nanoparticles (UCNPs) were simultaneously loaded with the photosensitizer chlorin e6 (Ce6) and the antibody glypican-3 (GPC3), which is overexpressed in hepatocellular carcinoma cells. The multitasking UCNPs@mSiO. We found that the UCNPs@mSiO. We believe that the utilization of 808 nm NIR excited UCNPs@mSiO

Topics: Animals; Antibodies; Cell Line, Tumor; Chlorophyllides; Glypicans; Hemolysis; Hep G2 Cells; Humans; Light; Liver Neoplasms; Male; Mice, Nude; Nanoparticles; Photochemotherapy; Photosensitizing Agents; Porphyrins; Xenograft Model Antitumor Assays

Theranostic nanoprobes integrated with dual-modal imaging and therapeutic functions, such as photodynamic therapy (PDT), have exhibited significant potency in cancer treatments due to their high imaging accuracy and non-invasive advantages for cancer elimination. However, biocompatibility and highly efficient accumulation of these nanoprobes in tumor are still unsatisfactory for clinical application. In this study, a photosensitizer -loaded magnetic nanobead with surface further coated with a layer of cancer cell membrane (SSAP-Ce6@CCM) was designed to improve the biocompatibility and cellular uptake and ultimately achieve enhanced MR/NIR fluorescence imaging and PDT efficacy. Compared with similar nanobeads without CCM coating, SSAP-Ce6@CCM showed significantly enhanced cellular uptake, as evidenced by Prussian blue staining, confocal laser scanning microscopy (CLSM) and flow cytometric analysis. Consequently, SSAP-Ce6@CCM displayed a more distinct MR/NIR imaging ability and more obvious photo-cytotoxicity towards cancer cells under 670 nm laser irradiation. Furthermore, the enhanced PDT effect benefited from the surface coating of cancer cell membrane was demonstrated in SMMC-7721 tumor-bearing mice through tumor growth observation and tumor tissue pathological examination. Therefore, this CCM-disguised nanobead that integrated the abilities of MR/NIR fluorescence dual-modal imaging and photodynamic therapy might be a promising theranostic platform for tumor treatment.

Topics: Adsorption; Animals; Biological Transport; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Membrane; Chlorophyllides; Humans; Infrared Rays; Lasers; Liver Neoplasms; Magnetite Nanoparticles; Mice; Mice, Nude; Optical Imaging; Photochemotherapy; Photosensitizing Agents; Porphyrins; Reactive Oxygen Species; Theranostic Nanomedicine; Tumor Burden; Xenograft Model Antitumor Assays

The sonodynamically induced anti-tumor effect of chlorin-e6 (Ce6) was studied in mice bearing hepatoma-22 solid tumors.. In order to determine the optimum timing of ultrasound exposure after administration of Ce6, the Ce6 concentrations in plasma, skin, muscle and tumor were estimated by measuring the fluorescence intensity of tissue extractions with a fluorescence photometer based on the standard curve. A three-dimensional optical imaging system (IVIS spectrum) was used further to characterize the distribution of Ce6 in H-22 tumor. The anti-tumor effects were estimated by measuring tumor size after sonodynamic therapy.. Similar pharmacokinetic trends of Ce6 in mice were observed either by fluorescence spectrophotometry or by bio-optical imaging. The results also demonstrated that Ce6 has a preferential localization in tumors, but low accumulation and rapid clearance in normal tissues. The results of anti-tumor effects revealed that at an ultrasound intensity of 4 W/cm(2) and a Ce6 dose of ≥10 mg/kg, a significant synergistic effect of ultrasound combined with Ce6 was observed, reducing the tumor volume significantly.. Chlorin-e6 is a potential sonosensitizer for fluorescence imaging as well as for sonodynamic therapy for cancer. The anti-tumor effect of ultrasound could be enhanced in the presence of Ce6, which might be involved in a sonochemical mechanism.

Topics: Animals; Antineoplastic Agents; Carcinoma, Hepatocellular; Cell Line, Tumor; Chlorophyllides; Drug Evaluation, Preclinical; Female; Fluorescence; Humans; Liver Neoplasms; Mice; Mice, Inbred ICR; Porphyrins; Radiation-Sensitizing Agents; Spectrometry, Fluorescence; Treatment Outcome; Ultrasonic Therapy; Xenograft Model Antitumor Assays

This paper provides the results of the non-clinical evaluation of biodistribution of the PS Photolon in inner organs and tissues of intact and tumor-bearing rats with xenograft tumors of different morphologic types.. The accumulation studies were performed in rats by means of intravital laser fluorimetry in situ and spectrophotometric determination ex vivo.. The biodistribution pattern of Photolon does not depend on tumor carriage as well as on morphologic type of the xenograft tumor. We have also showed that Photolon easily crosses an intact blood-brain barrier and accumulates in tissues of central nervous system. The relative bioavailability of brain tissues for Photolon was estimated as 82%, T(max)-30 min, mean residual time (MRT)-1.6h.. In general, results of the experimental study of biodistribution of Photolon in inner organs and tissues of rats, performed as in real time (by means of intravital laser fluorimetry in situ) as ex vivo (spectrophotometric assay) can be utilized while optimizing existing regimens of PDT with the purpose to increase safety and efficacy of treatment as well as for the development of new PDT protocols with Photolon applied for new indications. Parameters of pharmacokinetics and biodistribution of Photolon/Fotolon as well as its' ability to cross an intact blood-brain barrier, are optimal for the majority of modern clinical applications of PDT.

Topics: Animals; Carcinosarcoma; Chlorophyllides; Female; Humans; Liver Neoplasms; Male; Porphyrins; Povidone; Protoporphyrins; Rats; Tissue Distribution; Transplantation, Heterologous

Novel benzochloroporphyrin derivatives (BCPDs) were designed, synthesized, and characterized. In vitro dark cytotoxicity and photodynamic efficacy of BCPDs were evaluated by MTT assay on human hepatoma BEL-7402 cells. The experimental results showed that BCPDs 15, 16, 17, and 18 have strong long wavelength absorptions around 670 nm and exhibit significantly lower dark cytotoxicity than BPDMA and possess potent photocytotoxicity, IC50 values 1.32 microg/mL for 15, 0.26 microg/mL for 16, 0.47 microg/mL for 17 of 0.27 microg/mL for 18, and 0.23 microg/mL for BPDMA. Among them, BCPDs 16 and 18 are more effective and promising PDT photosensitizers based on the studies with BEL-7402 cells and show nearly the same photodynamic efficacy as BPDMA. MG-P staining qualitative analysis also indicated that PDT with BCPDs 16 can induce apoptosis in BEL7402 cells.

Topics: Carcinoma, Hepatocellular; Cell Line, Tumor; Chlorophyllides; Drug Design; Humans; Liver Neoplasms; Photochemotherapy; Photosensitizing Agents; Porphyrins; Protoporphyrins; Spectrophotometry, Ultraviolet

Photodynamic therapy has been applied quite extensively over the last few years, whereby the activation of photosensitizers by light causes the production of reactive oxygen species such as singlet oxygen, which is cytotoxic. The goal of this study was the enhancement of the photodynamic activity of photosensitizers through their delivery to specific, sensitive intracellular compartments of target cells. We synthesized a BSA-insulin-chlorin e6 conjugate that bound specifically to the insulin receptors (EC50, 1 nM) of the human hepatoma cell line PLC/PRF/5 and could be internalized by receptor-mediated endocytosis. Photodynamic activity, as assessed by various tests, indicated EC50s at about 100 times lower concentrations of conjugate compared to free chlorin e6 itself; and lower doses of irradiation were necessary to activate the conjugate compared to free chlorin e6. Inhibition of endocytosis of the conjugate abrogated the enhanced photodynamic activity of the conjugate above that of free chlorin e6. Endocytosis and subsequent localization around and in the cell nucleus of the BSA-insulin-chlorin e6 conjugate could be visualized using both FITC-labeled conjugate and 2',7'-dichlorofluorescin diacetate, a fluorescent indicator of the production of active oxygen species due to chlorin e6 activation. It was concluded that photodynamic activity of the conjugate is higher than that of free chlorin e6 through its receptor-mediated delivery into sensitive intracellular compartments.

Topics: Carcinoma, Hepatocellular; Chlorophyllides; Endocytosis; Humans; Insulin; Intracellular Fluid; Liver Neoplasms; Photochemotherapy; Porphyrins; Radiation-Sensitizing Agents; Serum Albumin, Bovine; Tumor Cells, Cultured

Transplantable rat tumours (sarcoma M-1, sarcoma 45, alveolar liver cancer PC-1 and Pliss' lymphosarcoma) were used to study chlorin e6 accumulation in tumours and its photodynamic effect. Tumours were irradiated by krypton ion laser light (647 and 676 nm; 90 J cm-2) 15 min and 24 h after chlorin e6 injection at doses of 5 and 10 mg kg-1. The relationship between some morphological peculiarities of these tumour strains, photosensitizer accumulation in tumours and their response to the photodynamic treatment is discussed.

Topics: Animals; Cell Division; Chlorophyllides; Female; Fibrosarcoma; Light; Liver Neoplasms; Lymphoma, Non-Hodgkin; Neoplasms, Experimental; Photochemotherapy; Porphyrins; Radiation-Sensitizing Agents; Rats; Rats, Wistar; Sarcoma, Experimental; Skin

Experiments with human hepatoma PLC/PRF/5 cells involving the use of two different tests (colony formation and Trypan blue exclusion) have demonstrated a significantly higher photosensitizing activity of chlorin e6 conjugates with bovine serum albumin (BSA) and internalizable ligand insulin as compared to that of chlorin e6 itself. Receptor-mediated internalization of insulin-BSA-chlorin e6 conjugates ensures greater photosensitization of cells than the binding of those conjugates to cell surface receptors. The suitability of such conjugates permitting the delivery of a photosensitizer to most sensitive cell structures is discussed.

Topics: Carcinoma, Hepatocellular; Chlorophyllides; Humans; Insulin; Liver Neoplasms; Photochemistry; Porphyrins; Radiation-Sensitizing Agents; Serum Albumin, Bovine; Tumor Cells, Cultured