Identification of the targets of hematoporphyrin derivative in lung adenocarcinoma using integrated network analysis

BACKGROUND: Hematoporphyrin derivative (HPD) has a sensibilization effect in lung adenocarcinoma. This study was conducted to identify the target genes of HPD in lung adenocarcinoma. METHODS: RNA sequencing was performed using the lung adenocarcinoma cell line A549 after no treatment or treatment with X-ray or X-ray + HPD. The differentially expressed genes (DEGs) were screened using Mfuzz package by noise-robust soft clustering analysis. Enrichment analysis was carried out using "BioCloud" online tool. Protein-protein interaction (PPI) network and module analyses were performed using Cytoscape software. Using WebGestalt tool and integrated transcription factor platform (ITFP), microRNA target and transcription factor (TF) target pairs were separately predicted. An integrated regulatory network was visualized with Cytoscape software. RESULTS: A total of 815 DEGs in the gene set G1 (continuously dysregulated genes along with changes in processing conditions [untreated-treated with X-ray-X-ray + treated with HPD]) and 464 DEGs in the gene set G2 (significantly dysregulated between X-ray + HPD-treated group and untreated/X-ray-treated group) were screened. The significant module identified from the PPI network for gene set G1 showed that ribosomal protein L3 (RPL3) gene could interact with heat shock protein 90 kDa alpha, class A member 1 (HSP90AA1). TFs AAA domain containing 2 (ATAD2) and protein inhibitor of activated STAT 1 (PIAS1) were separately predicted for the genes in gene set G1 and G2, respectively. In the integrated network for gene set G2, ubiquitin-specific peptidase 25 (USP25) was targeted by miR-200b, miR-200c, and miR-429. CONCLUSION: RPL3, HSP90AA1, ATAD2, and PIAS1 as well as USP25, which is targeted by miR-200b, miR-200c, and miR-429, may be the potential targets of HPD in lung adenocarcinoma.

Progression of basic research, clinical application of photodynamic therapy and fluorescence-guided surgery in glioma treatment / 中南大学学报(医学版)

The prognosis of glioma remains poor because of the infiltrative nature and the high local relapse rate.The current goals for patients with gliomas is maximal safe resection and adjuvant therapy.Tumor-specific photosensitizer,such as hematoporphyrin derivative (HpD) 5-aminolevulinic acid (5-ALA),can be selective up-taken and accumulated in tumor tissue.Light with appropriate wavelength can penetrate tumor tissue and excite the photosensitizer.The excited photosensitizer within glioma cells permits fluorescence visualization of tumor tissue during surgery and has been introduced in treatment of glioma as fluorescence-guided surgery (FGS).On the other hand,the toxicity of singlet oxygen generated by the excited photosensitizer has been used as photodynamic therapy (PDT) in selective destruction of the tumor.Some reports demonstrated the usefulness of adding PDT as an intraoperative adjuvant therapy,but the complexity associated with its implementation and the introduction of TMZ prevented PDT from becoming a routine therapy.However,FGS using 5-ALA in patients with malignant brain tumors has surfaced globally and may become a useful tool in increasing the extent of resection in gliomas.

Investigating the killing effect of photodynamic therapy induced by different photosensitizers on human leukemia cell in vitro / 国际生物医学工程杂志

Objective To investigate and compare the killing effect of photodynamic therapy (PDT)induced by hematoporphyrin derivative (HpD),hematoporphyrin monomethyl ether (HMME) and photocarcinorin (PsD007) on human leukemia cells K562 in vitro.Methods Human leukemia cells were cultured with serial concentrations of photosensitizers followed by irradiation of different dosage of laser light,then MTT colorimetric assay was applied to measure the relative survival rate of PDT for the cells.Results Significant difference in the inhibitory between the PDT group and control group was observed (P＜0.05).The survival rate of PDT for the cells elevated along with the increase in the concentration of sensitizer and dose of laser light.When the photosensitizer concentration was bigger (25 μg/ml) or the energy density was bigger (7.2 J/cm2),the effect of PsD007 was better than HMME,and they were significantly better than HpD (P＜0.05).Conclusion PDT has significant killing effect on human leukemia cells K562,and its relative inhibitory rate appears to be correlated with the dose of sensitizer and laser light irritation.The effect of PDT is related to the photosensitizers.The effect of HpD-PDT is not as effective as PsD007 and HMME.On the conditions of higher energy density and larger photosensitizer concentration,the effect of PsD007-PDT is better than HMME-PDT.

Photodynamic inactivation of four Candida species induced by photogem®

This study evaluated the in vitro susceptibility of C. albicans, C. dubliniensis, C. tropicalis and C. krusei to photodynamic therapy (PDT) induced by Photogem® and light emitting diode (LED). Suspensions of each Candida strain were treated with three photosensitizer (PS) concentrations (10, 25 and 50 mg/L) and exposed to 18.0, 25.5 and 37.5 J/cm² LED light fluences (λ ~ 455 nm). Control suspensions were treated only with PS concentrations, only exposed to the LED light fluences or not exposed to LED light or PS. Sixteen experimental conditions were obtained and each condition was repeated three times. From each sample, serial dilutions were obtained and aliquots were plated on Sabouraud Dextrose Agar. After incubation of plates (37 ºC for 48 hours), colonies were counted (cfu/mL) and the data were statistically analyzed by ANOVA and the Tukey test (α=0.05). Complete killing of C. albicans was observed after 18.0 J/cm² in association with 50 mg/L of PS. C. dubliniensis were inactivated after 18.0 J/cm² using 25 mg/L of PS. The inactivation of C. tropicalis was observed after photosensitization with 25 mg/L and subsequent illumination at 25.5 J/cm². For C. krusei, none of the associations between PS and light resulted in complete killing of this species. PDT proved to be effective for the inactivation of C. albicans, C. dubliniensis and C. tropicalis. In addition, reduction in the viability of C. krusei was achieved with some of the PS and light associations.

Hematoporphyrin derivative photodynamic therapy of human pancreatic cancer cells in vitro / 中华胰腺病杂志

Objective To investigate the killing effect of hematoporphyrin derivative photedynamic therapy (PDT) on cultured human pancreatic cancer cell,and to explore the mechanism of this effect.Methods Biolitec PDT 630 semi-conductor laser therapeutic apparatus was used as the light source.After pancreatic cancer cell PANC1 was incubated 8 h with different concentrations of Photosan(hematoporphyrin derivative) as photosensitizer (0.5mg/L,1 mg/L,2 mg/L,4 mg/L),the cells were given different doses of 630nm laser irradiation(1 J/cm2' 5 J/cm~2,10 J/cm~2 ).The A492 value was determined in each group with MTT method.Cell apoptosis rate was detected by flow cytometry after PDT.Results There was no killing effect when no Photosan was administrated;10 J/cm~2 irradiation had killing effect on PANC1 when Photosan was administrated as 1 mg/L(0.140±0.013 vs 0.213±0.008,P＜0.05);5 and 10 J/cm~2 irradiation all had killing effect on PANC1 when Photosan was administrated as 2 mg/L (0.081±0.024 and 0.049±0.013vs 0.211±0.031,P＜0.05 and P＜0.01 );all doses of irradiation had killing effect when Photosan was administrated as 4 mg/L.There was no significant difference between 5 and 10 J/cm~2 irradiation in term of killing effect.Cell apoptosis rates with 0 or 2 or 4 mg/L Photosan and 10 J/cm~2 irradiation were(13.8±1.8) %,(40.9±1.6)%,(62.5±2.0)%,the difference was statistically significant(P＜0.05).Conclusions Photosensitizer or irradiation alone did not produce PDT effect.With certain dose of photosensitizer and irradiation,the PDT effect increased accordingly.

Experimental interstitial photodynamic therapy of the Morris 7777 hepatoma in the rat / 대한내과학회지

BACKGROUND: Photodynamic therapy (PDT) of hepatic tumors has been restricted due to the preferential retention of photosensitizers in normal liver tissue. Therefore, superficial tumor illumination causes subsequential liver necrosis. Moreover, the limited light penetration during superficial illumination makes it impossible to treat deep-seated or larger solid tumors. These limitations were overcome by interstitial therapy. Aim: The aim of study is to investigate the macroscopic and microscopic effect of a single session of interstitial photodynamic therapy on liver tumor. METHODS: The Morris 7777 hepatoma cells were injected subcutaneously into the flanks of Buffalo female rats. Tumor growth was monitored with measuring the tumor size. Animals were pretreated with hematoporphyrin derivative 48 hours prior to the intratumoral delivery of laser radiation, when the tumor had reached a volume greater than 1.0 cm3. One or two weeks after interstitial PDT, the extent of tumor necrosis was evaluated microscopically. RESULTS: Volume of lesions averaged 10.2 mm3. Histologic stains demonstrated microvascular thrombosis and coagulative necrosis within the lesions. There appeared to be 100% cellular destruction within the lesion by cytochemical staining. CONCLUSION: The results of this study show the ability of interstitial PDT to cause destruction of liver tumor.

The study of photodynamic effect induced by laser on human tumor cell lines and umbilical cord MNC in vitro / 中华物理医学与康复杂志

Objective To investigate the photodynamic effect induced by laser and use it as a method for purging of minimal residual tumor cells from autologous peripheral blood stem cell grafts. Methods We studied the effect of various concentrations of hematoporphyrin derivative (HPD) combined with of different time of laser irradiation on human leukemia cell line K562, breast tumor cell line MCF-7 and human umbilical cord blood-derived hematopoietic stem/progenitor cells by using MTT assay to compare the photodynamic effect between cell lines of each group. Results HPD or laser irradiation alone had no apparent cytotoxicity while the application of HPD plus laser exposure caused photosensitization. HPD plus laser irradiation was more efficient for killing K562 cells and MCF-7 cells than doing the normal human umbilical cord blood-derived hematopoietic stem/progenitor cells. The photodynamic effect on K562 cells and MCF-7 cells was positively correlated with the increase of treatment duration and HPD concentration. Conclusion These results indicated that the laser photodynamic effect mediated by HPD might be used for purging autologous peripheral blood stem cell grafts in vitro.

Comparative study on the killing effect between hypocrellins B-photodynamic therapy and hematoporphyrin derivative-photodynamic therapy on human lung cancer cells / 解放军医学杂志

Objective To study the killing effect of hypocrellins B-photodynamic therapy (HB-PDT) for lung cancer cell line A549, to compare with that of hematoporphyrin derivative-photodynamic therapy (HpD -PDT), in order to define the superiority of HB-PDT in the therapy of lung cancer. Methods Lung cancer cell line A549 was used in the study. The cells were incubated in vitro with HB or HpD dissolved in DMEM in different concentrations, and then irradiated by copper vapor laser with mixed wavelength light under saturated light dose. Cell survival rate was respectively measured by MTT assay after 24 hours' incubation. According to cell survival curves after being cultured with different photosensitizers in different concentrations, the equation of each cell survival curve was plotted. Finally IC 50 (50% inhibition concentration) of each photosensitizer was derived. Results The results showed that HB-PDT had a strong killing effect on lung cancer cell. The IC 50 of HB was 33.82ng/ml for lung cancer cell, while the IC 50 of HpD was 1 316.88ng/ml, which was 38.94 fold of that of HB. Conclusions HB was a more effective photosensitizer than the photosensitizer HpD. HB-PDT had strong photodynamic killing effect on lung cancer cell line.

Distribution of Hematoporphyrin Monomethyl Ether in Tumor-bearing Mice / 第二军医大学学报

spleen(?)heart(?)muscle.The pharmacokinetics of PsD-044 in mice was fitted by an open three compartment model. The pharmacokinetic parameters calculated were shown to be essentially consistent with the determination of tissue distribution. It was also shown that PsD-044 can't pass through blood-brain barrier.

PATHOLOGICAL OBSERVATIONS ON ACUTE AND SUBACUTE TOXICITY OF PHOTOCARCINORIN(PsD-OO7)IN DOGS / 第二军医大学学报

Preclinical toxicology of PsD-007(a new photosensitizer prepared in our coll-ege)was studied.Pathological observations were performed on major organs of the toxic dogs including liver, kidneys, myocardium, lungs, spleen, stomach, intestine, mesenteric lymph nodes, urinary bladder, prostata, testes, ovary, adrenals, pituitary gland, thyroid and brain.Focal balloning degeneration of hepatic cells occurred in lower dose groups, and degeneration and necrosis of hepatocytes and proximal convoluted tubular epithelial cells of kidney, and edema of brain in higher dose groups were observed in acute toxicity testing.The hepatocytes were normal, but hyperplasia, swelling and vacuoles within the cytoplasm of the Kupffers cells in liver, necrosis of tubules of kidney and edema of brain in higher dose groups were found in subacute testing. These damages were reversible within 4 weeks except in higher dose groups, and their morphologic severity was dose-related.The results show that the liver and kidney may be the main target organs of toxicity of PsD, and the detections of hepatic and renal functions may be useful to guard against its toxic effects during clinical use.

STUDY ON CLINICAL PHARMACOKINETICS OF THE NEW PHO- TOSENSITSZER PHOTOCARCINORIN FOR TUMOR LOCALIZATION AND PHOTORADIATION THERAPY / 第二军医大学学报

The clinical pharmacokinetic behavior of new photosensitizer photocarcinorin for the photoradiation therapy of 11 patients with different malignant tumors was studied by means of fluorospectrophotometric method after intravenous infusion.The dose infused was 4-4.5 mg/kg body weight and the infusion rate, 0.1 mg/ kg/min.It is a three-compartment model as determined by the drug-time curve through Akaike's Information Criterion(AIC) .The principal pharmacokinetic parameters calculated by nonlinear least-square regression program are: t1/2? = 0.26?0.05h, t1/2? = 4.52?1.04h, t1/2?=32.9?5.31h.In addition, the pharmacokinetic parameters of one patient injected with lower dose(1.82 mg/kg)were also determined, and shown as a three compartment model.

STUDIES ON THE CHEMISTRY OF PHOTOCARCINQRIN / 第二军医大学学报

The preparation,chemical composition and physicochemical properties of photocar-cinorin (PsD-007),a new drug of photolocalizaion and photochemotherapy for human malignancies,are reported here.It has been shown by the resul s of thin layer and high performance liquid chromatographic (TLC & HPLC) analyses that about 85% of its total weight are relatively hydrophobic porphyrins of unknown structure in which a fraction with the same retention time as .hematoporphyrin(Hp)monoacetate constituts over 60%.There are also 4.2% of Hp,8.6% of 3(8) hydroxycthy lS-(3)-vinyldcutero-porphyrin (HVD) and 0.6% of portoporphyrin (Pp) in this drug.The presence and amount of HVD in PsD-007 were determined by an authentic sample prepared in our laboratory.The changes of the composition of PsD-007 on exposure to different doses of argon laser (488+ 514.5nm) were detected by HPLC.The variations in Soret peak of UV absorption spectra of PsD-007 protected from light at a constant temperature were also determined

EFFECT OF ULTRASONICALLY ACTIVATED HEMATOPORPHYRIN ON EHRLICH ASCITES TUMOR CELLS IN VITRO / 解剖学报

Objective The aim of this study was to investigate the effect of ultrasonically activated hematoporphyrin on ultrastructure of ehrlich ascites tumor(EAT) cells and to evaluate the potential mechanism of action inducing this cytotoxicity. Methods EAT cells in vitro were exposed to ultrasound at 2

THE UPTAKE AND RETENTION OF ~3H-HEMATOPORPHYRIN DERIVATIVE IN HUMAN MALIGNANT AND NORMAL CELLS IN VITRO / 解剖学报

The uptake and retention of ~3H-HPD in.human malignant cells(cell line MGC 80-3,SGC-7901,BEL-7402,Os-732,Detroit-6 and HeLa)and normal fibroblasts were studied by liquid scintillation counting method.The order of cell lines which took up ~3H-HPD after 30 min.incubation were fibroblasts,HeLa, BEL-7402,Detroit-6,SGC-7901,MGC 80-3 and Os-732 respectively according to the amount of ~3H-HPD in cells.Due to the variety in the amount of excretion, however,those cells which retained ~3H-HPD after 24 hours cultivation were Os-732, Detroit-6, BEL-7402,HeLa,SGC-7901,MGC 80-3 and fibroblasts respectively in respect to the ~3H-HPD level in cells.The uptake was reduced by raising extracel- lular pH and lowering temperature as well as adding higher concentration of serum. Our results showed that the ~3H-HPD content in malignant ceils was higher than those in normal ones and it was because that malignant cells retained ~3H-HPD longer other than took up more

EFFECTS OF HEMATOPORPHYRTN DERIVATIVE (HpD)-LASER ON THE KIDNEYS OF NORMAL MICE——OBSERVATION WITH LIGHT MICROSCOPY ELECTRON MICROSCOPY AND HISTOCHEMISTRY / 解剖学报

Fortyfour male Kunming mice, weighing 20-25g. were used. The HpD was injected (10mg/kg) into the caudal vein of 32 mice individually. After 48 hours, the kidneys were radiated directly by the dye-laser for 10 minutes (630nm, 250-300J/cm~2). Then, the kidneys were observed at different times. 12 normal male mice served as control. The experiment demonstrated that when HpD was activated by laser, it could produce the necrotic effect on the renal tissues. The different structures of the kidney showed different susceptibility to HpD-laser. The proximal convoluted tubules and capillaries were injured earlier than the distal convoluted tubules and the collecting tubules. The first morphologic changes observed electron microscopically were the swelling of mitochondria and the formation of vesicles on the cell membrane. Finally, they were completely disrupted. The damages of lysosomes, endoplasmic reticula, ribosomes and cell nucleus followed. The activity of some enzymes in the kidney were inhibited. The sequence of the extents of the enzymes being inhibited was AkP, AcP, Cytox, SDH. The mucopolysaccharides ran off the brush border in proximal convoluted tubules. The results of this study suggested that the various susceptibilities to HpD-laser are chiefly due to the different structures and their functional conditions in the kidney. The main photodynamic effect is presented as the injury of the membrane system of the cell.

PHOTODYNAMIC EFFECT OF HEMATOPORPHYRIN DERIVATIVE (HPD) ON MITOCHONDRIA AND SUCCINATE DEHYDROGENASE OF HUMAN GASTRIC CANCER CELLS IN DIFFERENT PHASES OF CELL CYCEL / 解剖学报

Synchronous cells in different phase (G_1, S, G_2, M) of cell cycle were obtained from human gastric low-differentiated mucous adenocarcinoma cells (MGC 80-3), which werec ultured with nitrous oxide under high pressure and blocked with overdosage of TdR. Dynamic changes of the mitochondria stained with Rhodamine-123 and succinate dehydrogenase demonstrated by cytochemical method were observed in various phase of the cells at 0, 24, 36, 60 hours after treatment with HPD plus red light. The results showed that mitochonodria of all four phases are of impairment immediately after photoradiation, SDH reactivity is decreased slightly at 24 hours, the activities of SDH is the weakest. As time goes on, we observed that mitochondria gradually recovered to its original structure and then SDH returned to its normal level. The recovery rate of mitochondria and SDH was in the following order, i. e. S, G_1, G_2, andM. The relationship between these changes and cell killing is briefly discussed.