CN115814080B - Photodynamic therapeutic agent containing cryptotanshinone and application thereof - Google Patents
Photodynamic therapeutic agent containing cryptotanshinone and application thereof Download PDFInfo
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Abstract
The invention discloses a photodynamic therapeutic agent containing cryptotanshinone and application thereof, belonging to the technical field of antitumor drugs, wherein the cryptotanshinone can induce active oxygen through photosensitization under the action of 460nm absorption light 1 O 2 Compared with the effect of the cryptotanshinone alone, the cryptotanshinone photosensitization can obviously reduce the survival rate of MDA-MB-231 cells and inhibit the migration of MDA-MB-231 cells. After MDA-MB-231 cells and cryptotanshinone act for 4 hours, the engulfment of the cryptotanshinone by the MDA-MB-231 cells reaches saturation, and when 460nm light absorption acts on the MDA-MB-231 cells containing the cryptotanshinone, the cryptotanshinone photosensitization can increase the intracellular ROS level, reduce the mitochondrial membrane potential of the cells, cause the MDA-MB-231 cells to generate G2/M blocking, and obviously induce apoptosis necrosis of the MDA-MB-231 cells.
Description
Technical Field
The invention belongs to the technical field of antitumor drugs, and in particular relates to a photodynamic therapeutic agent containing cryptotanshinone and application thereof.
Background
Cryptotanshinone is a phenanthrenequinone compound extracted from traditional Chinese medicine radix salviae miltiorrhizae-Salvia miltiorrhiza Bunge. Cryptotanshinone as an important effective component of Saviae Miltiorrhizae radix has various pharmacological activities including antiinflammatory, antibacterial, antioxidant and anti-obesity, and also has good anticancer effect. The cryptotanshinone has proliferation inhibiting effect on various cancer cells, including colorectal cancer, breast cancer, prostatic cancer, cervical cancer, rhabdomyosarcoma, melanoma, etc. Chemosensitization drugs are used to increase the sensitivity of NQO 1-highly expressing malignant tumor cells to anticancer drugs. The patent CN202210731322.8 of 26 9 months of 2022 discloses application of combination of cryptotanshinone and NQO1 activating medicament in preparing a chemotherapeutic sensitization medicament for malignant tumor with high NQO1 expression, and aims to improve the killing effect of the NQO1 activating medicament on malignant tumor cells, and present Coalism and synergistic effects, so that the dosage of the NQO1 activating medicament is obviously reduced, and the problem that the NQO1 activating medicament has a plurality of clinical practical application problems of low efficacy, insensitivity or medicament tolerance, poor curative effect caused by relapse, strong toxic and side effects, treatment failure and the like of the NQO1 activating medicament for treating or preventing tumor at present is solved. Breast cancer is one of the most common malignant tumors of females, the incidence rate of which increases year by year, and the death rate of female cancers is the first. Traditional methods of treating cancer include surgical excision therapy, radiation therapy, and chemotherapy, which are traumatic to the human body, have many postoperative complications, and are prone to metastasis of tumor cells. Radiation therapy has obvious physiological toxicity to human body, while chemotherapy principle produces great damage to normal cells of human body. Wherein, estrogen, progestogen and HER-2 are used for expressing negative triple negative breast cancer-triple negative breast cancer-TNBC, and the malignant degree is highest, and the survival rate is poorer than that of non-triple negative breast cancer because of endocrine treatment and anti-HER-2 targeted treatment. Zhou Nayang and the like show that the cryptotanshinone can obviously inhibit the activity, migration and invasion of the MDA-MB-231 cells of the triple negative breast cancer in vitro, but is dose-dependent. New drug studies aimed at combating triple negative breast cancer are one of the key points of current breast cancer treatments. As those skilled in the art, development of a photodynamic therapeutic agent containing cryptotanshinone and its use is desired.
Disclosure of Invention
The invention aims to provide a photodynamic therapeutic agent containing cryptotanshinone and application thereof, so as to solve the technical problems.
A photodynamic therapeutic agent comprising cryptotanshinone, the active ingredient of which comprises cryptotanshinone.
The photodynamic therapeutic agent containing the cryptotanshinone is applied to preparing the medicines for preventing, relieving and/or treating triple negative breast cancer.
The use method of the photodynamic therapeutic agent containing the cryptotanshinone comprises the following steps: cryptotanshinone is used as photosensitizer and activated by light.
Further, the photoactivation comprises long wavelength light, wherein the long wavelength light comprises wavelengths between 460 and 520 nm.
Further, after the photo-activation of cryptotanshinone, free radical oxygen species with cytotoxicity are released.
The cryptotanshinone serving as a photosensitizer is activated by light to reduce the survival rate of MDA-MB-231 cells and inhibit the migration of MDA-MB-231 cells.
The active ingredient of the photodynamic therapeutic agent comprises cryptotanshinone;
further, it is preferred that the pharmaceutical carrier is included in a variety of pharmaceutically acceptable formulations and one or more pharmaceutically acceptable excipients.
Cryptotanshinone, CAS number: 35825-57-1, cryptotanshinone, hereinafter CTan, has the chemical structural formula:
the invention is realized in the following way:
in a first aspect, the present invention demonstrates for the first time that cryptotanshinone, CTan, an effective photosensitizer capable of inducing reactive oxygen species by photosensitization upon absorption of light at 460nm 1 O 2 Is generated.
In a second aspect, the invention provides a new idea and method for applying CTan as photosensitizer to photodynamic anti-breast cancer DA-MB-231 cells in the anti-tumor field.
In a third aspect, CTan photosensitization significantly reduces MDA-MB-231 cell viability and inhibits MDA-MB-231 cell migration as compared to CTan effect alone. Meanwhile, the invention discusses the related mechanism, and the result shows that after MDA-MB-231 cells act with CTan for 4 hours, the phagocytosis of CTan by the MDA-MB-231 cells reaches saturation;
in the fourth aspect, when 460nm light absorption is performed on MDA-MB-231 cells containing CTan, CTan photosensitization can increase intracellular ROS level, reduce mitochondrial membrane potential of the cells, cause the MDA-MB-231 cells to generate G2/M retardation, and obviously induce apoptosis necrosis of the MDA-MB-231 cells.
In a fifth aspect, the invention also provides a photodynamic therapeutic agent for resisting breast cancer MDA-MB-231 cells, an active ingredient of the photodynamic therapeutic agent comprises cryptotanshinone, and application of the photodynamic therapeutic agent in preparing a medicament for preventing, relieving and/or treating triple negative breast cancer.
MTT, chemical name 3- (4, 5-dimethylthiazole-2) -2, 5-diphenyltetrazolium bromide, trade name: thiazole blue. The detection principle is that succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-violet crystalline Formazan (Formazan) and deposit in cells, whereas dead cells do not. Dimethyl sulfoxide (DMSO) can dissolve formazan in cells, and the light absorption value of the formazan can be measured at 490nm wavelength by an enzyme-labeled instrument, and the MTT crystallization forming amount is proportional to the cell number in a certain cell number range. Based on the measured absorbance value (OD value), the number of living cells was judged.
The invention has the following beneficial effects:
the invention for the first time proves that the cryptotanshinone (CTan) is an effective photosensitizer which can induce active oxygen through photosensitization under the action of 460nm absorption light 1 O 2 Is generated. The present invention thus innovatively utilizes CTan as a photosensitizer for the study of photodynamic anti-breast cancer MDA-MB-231 cells. The results indicate that CTan photosensitization significantly reduces the survival of MDA-MB-231 cells and inhibits migration of MDA-MB-231 cells as compared to CTan effect alone. Meanwhile, the invention discusses related mechanisms, and results show that when MDA-MB-231 cells and CTan act for 4 hours, the phagocytosis of the MDA-MB-231 cells to the CTan reaches saturation, and when 460nm light absorption acts on the MDA-MB-231 cells containing the CTan, the CTan photosensitization can increase intracellular ROS level, reduce the mitochondrial membrane potential of the cells, cause the MDA-MB-231 cells to generate G2/M retardation, and obviously induce apoptosis necrosis of the MDA-MB-231 cells.
The invention provides a new idea and method for the application of CTan in the anti-tumor field.
Drawings
FIG. 1 is a graph showing the effect of MTT assay cryptotanshinone (CTan) photosensitization on breast cancer MDA-MB-231 cell viability, wherein CTan concentrations of 1, 1.5, 1.75, 2, 2.5, 3, and 3.5 μg/mL were treated, incubated for 4 hours, then irradiated with a 30W LED lamp panel having a center wavelength of 460nm for 30 minutes, then cultured for 24 and 48 hours, respectively, for MTT assay; FIG. 2 is a photograph of a cell scratch repair assay (wound-sizing assay) analyzing the effect of CTan photosensitization on MDA-MB-231 migration ability; FIG. 3 (A) is a comparison of CTan and DPBF UV-visible absorption spectra and structural formula; FIG. 3 (B) is a graph showing the change in DPBF content with irradiation time by high performance liquid chromatography. FIG. 3 (C) is a graph showing the change in DPBF content in the sample by high performance liquid chromatography. FIG. 4 is a graph of the incubation time of drug with cells versus the fluorescence intensity of CTan in MDA-MB-231 cells, wherein (A) is a superimposed visual graph and FIG. 4 (B) shows the fluorescence intensity of samples at different incubation times; FIG. 5 is a graph of the effect of CTan photosensitization on apoptosis and necrosis of MDA-MB-231 cells; FIG. 6 is a graph of the effect of CTan photosensitization on intracellular ROS levels in MDA-MB-231 cells. MDA-MB-231 cells were incubated with 2. Mu.g/mL CTan for 4 hours, then irradiated with 30W LED lamp panel having a center wavelength of 460nm for 30 minutes, and the cells were treated after the irradiation was completed for flow cytometry analysis. Wherein (A) a control group, (B) an illumination group alone, (C) a drug group alone, and (D) a drug and illumination act together; FIG. 7 is a graph of changes in mitochondrial membrane potential of CTan photosensitizing mediated MDA-MB-231 cells, wherein (A) is a control group, (B) is an illumination-alone group, (C) is an cryptotanshinone-alone group (2. Mu.g/mL), and (D) is a CTan-and-illumination synergy group (CTan concentration of 2. Mu.g/mL); FIG. 8 is a graph showing the effect of CTan photosensitization on MDA-MB-231 cell cycle distribution.
Detailed Description
The invention is illustrated, but not limited, by the following specific examples.
The following equipment and raw materials were used: high performance liquid chromatograph (Shimadzu LC-20 AB), cryptotanshinone (Ala Ding Shenghua), fetal bovine serum (Sijiqing), pancreatin digestive juice (Biyun Tian); annexin V-FITC/PI apoptosis detection kit, active oxygen detection kit (mitochondrial membrane potential detection kit JC-1) and flow cytometer (Beckman MoFlo XDP), cell cycle and apoptosis detection kit, laser scanning confocal microscope Zeiss LSM710, ultraviolet-visible spectrophotometer (T6-1650E). Breast cancer MDA-MB-231 cells at 37℃with 5% CO 2 Under the condition, the culture is routinely carried out on RPMI1640 medium containing 10% of fetal bovine serum. When the cells grow to about 80% of the monolayer, digestion and passage are carried out by using 0.25% trypsin solution containing 0.02% EDTA, and the cells with good growth are taken for experiment. MDA-MB-231 cells were incubated with 2. Mu.g/mL CTan for 4 hours, specifically with 1X 10 MDA-MB-231 cell suspension 5 Density of wells/plates in 6-well plates, cells were placed in 5% CO 2 The cells were incubated in an incubator at 37℃for 24 hours and then with 2. Mu.g/mL CTan for 4 hours.
Example 1
After cells were treated with different concentrations of CTan (1, 1.5, 1.75, 2, 2.5, 3 and 3.5. Mu.g/mL), incubated for 4 hours, then irradiated with a 30WLED lamp plate having a center wavelength of 460nm for 30min, then cultured continuously for 24 and 48 hours, respectively, for MTT analysis, each sample was repeated three times.
Cell activity was measured by MTT assay: MDA-MB-231 cells were inoculated into 96-well plates, 100. Mu.L of cell suspension was added to each well, and the cell count was 1X 10 4 Holes, 5 multiple holes per group. After 24h of culture, the original culture solution is absorbed, incubated for 4 hours by CTan treatment with different concentrations, then irradiated for 30min by a 30WLED lamp panel with the center wavelength of 460nm, and then cultured continuously, and the absorbance value of each group of samples is detected at the wavelength of 490nm by an MTT method (A490). Cell viability = (experimental group a value-control group a value)/(control group a value-blank group a value) ×100%, control group defined as 100%.
As can be seen from fig. 1: MTT results at 24 hours and 48 hours indicate that the cryptotanshinone (CTan) effect alone can produce a dose-dependent inhibitory effect on breast cancer MDA-MB-231 cell viability in the non-light condition, because CTan itself has a certain anticancer effect, but CTan alone has limited effect, and the survival rate of MDA-MB-231 cells is above 60% in all tanshinone IIA alone groups. However, when light and CTan combined, the survival of MDA-MB-231 cells was significantly reduced compared to the corresponding non-light group. This result indicates that illumination can significantly increase the toxic effect of CTan on MDA-MB-231 cells.
Example 2
Cell scratch repair assay (wound-sizing assay): the effect of CTan photosensitization on MDA-MB-231 migration ability was analyzed. MDA-MB-231 cells were scratched after cell monolayers were formed by adherence to a 6-well plate, then the cells were incubated with 2. Mu.g/mLCTan for 4 hours, then irradiated with a 30WLED lamp plate having a center wavelength of 460nm for 30 minutes, and after the irradiation was completed, the cells were continuously cultured, and after culturing for 0 and 24 hours, respectively, were photographed using a microscope.
As can be seen from fig. 2: the cell scratch repair test (work-sealing assay) is a test for examining the in-vitro migration capability of cells, which is low in cost, simple and easy to operate. The invention utilizes the Wound-sizing assay to examine the influence of cryptotanshinone (CTan) photosensitization on the migration of breast cancer MDA-MB-231 cells. In the control group, cell scratches were almost repaired as the incubation time reached 24 hours, and the illumination alone had little effect on cell migration compared to the control group, so that the repair degree of cell scratches was the same as that of the control group at 24 hours of cell culture. The CTan effect alone inhibited the migration ability of the cells to some extent, and after 24 hours of cell culture, the presence of cell-free areas was still visible although the scratches of the cell monolayer were repaired. However, when light and CTan are cooperated, after 24 hours of culture, cell-free areas formed by cell scratches are hardly repaired, so that the experiment shows that the photodynamic effect of CTan effectively inhibits the migration capacity of MDA-MB-231 cells.
Example 3
High performance liquid chromatography analysis of the change in DPBF content with irradiation time: the samples were DMSO solutions containing DPBF (1 mM) and CTan (0 or 1 mM) and were irradiated for different times (0, 10, 20, 30, 40, 50, 60, 70 and 80 min) under the action of light having a center wavelength of 520nmLED at 30W. (2) High performance liquid chromatography analysis of changes in DPBF content in samples: the sample is DMSO solution containing DPBF (1 mM) and CTan (1 mM), and is obtained by respectively ventilating for 20min by argon, air and oxygen, and irradiating for 30min under the action of light with a 30W central wavelength of 520 nmLED.
As can be seen from fig. 3: 1 O 2 can generate irreversible reaction with DPBF to cause photodegradation of DPBF, and the degradation of DPBF is analyzed in the practical experimental process to evaluate 1 O 2 Is generated. According to the invention, the change of the DPBF content is detected by adopting a high performance liquid chromatograph, as shown in FIG. 3A, the DPBF is not light-absorbed in a wavelength range larger than 480nm, in theory, the light emitted by the LED lamp panel with the center wavelength at 520nm can only be absorbed by CTan, and the direct photodegradation of the DPBF can be avoided. In the absence of CTan, the DPBF content still decreases with increasing illumination time, as shown in FIG. 3B, which illustrates that the selected light source may emit a portion of light that can be absorbed by the DPBF, resulting in direct photodegradation of the DPBF. However, the presence of CTan can significantly reduce the DPBF content at the same illumination time. The method comprisesSome experimental results show that CTan photosensitization produces 1 O 2 . In this experiment, DPBF is not light-absorbing in the wavelength range of more than 480nm, so most of light emitted by the LED lamp should be absorbed by CTan (although partial light can be absorbed by DPBF to cause degradation of DPBF), so as to cause CTan light excitation to generate corresponding CTan triplet excited state [ ] 3 CTan). At O 2 In the presence of the catalyst, 3 TanIIA can be replaced by O 2 Rapid quench generation 1 O 2 ， 1 O 2 Then reacts with DPBF to cause a great reduction in DPBF content.
O 2 Is a quencher with effective triplet excitation state, and the excited triplet state of the photosensitizer can be recovered to the ground state through attenuation of a non-radiative form, and can also be transferred to O through energy transfer 2 ，O 2 In the ground state, is in a triplet state and forms when receiving the energy of the excited triplet state of the photosensitizer 1 O 2 ，O 2 Content of (2) is to 1 O 2 The generation plays a vital role. For this purpose the invention examines O 2 Concentration differential pair 1 O 2 The effect of the generation. Firstly preparing a DMSO solution containing DPBF (1 mM) and CTan (0 or 1 mM), and respectively using argon, air and oxygen to ventilate and saturate the solution system so as to prepare three reaction systems with different oxygen concentrations. As shown in FIG. 3C, under the same irradiation conditions, the DPBF photodegradation by CTan photosensitization is O 2 Concentration-dependent, i.e. DPBF photodegradation with O 2 The increase in concentration increases. This can be explained by O 2 The increase in concentration can promote O 2 And 3CTan, and further increases 1 O 2 The amount produced eventually causes rapid degradation of the DPBF. In summary, CTan photosensitization can generate 1 O 2 ， 1 O 2 The generation of (c) provides theoretical support for the use of CTan as a photosensitizer.
Example 4
Flow cytometry analyzed phagocytosis of cryptotanshinone (CTan) by breast cancer MDA-MB-231 cells:
after breast cancer MDA-MB-231 cells were reacted with 2. Mu.g/mL of CTan for 0h, 1h, 2h, 4h, 6h and 8h, respectively, fluorescence emission intensity of CTan was detected on a flow cytometer using FITC analysis channels (each sample was repeated three times). Wherein (A) is a superposition visual chart, and (B) is fluorescence intensity of different samples.
As can be seen from fig. 4: prior to conducting extensive cell level experiments, the present invention examined the phagocytosis of CTan by breast cancer MDA-MB-231 cells to assess the ability of the drug to enter the cell, as well as the possible distribution within the cell. Analysis was performed herein by detecting CTan autofluorescence using a flow cytometer and confocal laser scanning microscope, respectively. We have found that under excitation with 405nm excitation light, CTan can emit fluorescence, and the wavelength range of the emission spectrum lies in the detection range of emission light of the flow cytometer, so that the flow cytometer can be used to directly detect CTan fluorescence to characterize the phagocytosis of CTan by MDA-MB-231 cells. As shown in fig. 4, the fluorescence intensity of CTan in MDA-MB-231 cells was gradually increased as the incubation time of the drug with cells was prolonged (from 1 hour to 8 hours), but the increase in fluorescence intensity was slowed down even within the error range when the drug was incubated with cells for more than 4 hours, so that it can be seen that the phagocytosis of CTan by MDA-MB-231 cells was close to saturation after the drug was allowed to act with cells for 4 hours, and thus this data also provides a reference for the subsequent cell experiments, i.e., the light experiments were performed after 4 hours after the drug was added, when the distribution of CTan in MDA-MB-231 cells was close to saturation.
Example 5
Effects of CTan photosensitization on apoptosis and necrosis of MDA-MB-231 cells:
MDA-MB-231 cells were incubated with 2. Mu.g/mLCTan for 4 hours, then irradiated with a 30WLED lamp plate having a center wavelength of 460nm for 30 minutes, and after the irradiation was completed, the cells were cultured for 24 hours, and then the treated cells were collected and analyzed using a flow cytometer.
As can be seen from fig. 5: the photodynamic effects of CTan effectively inhibit cell survival and migration, and the invention next examined the death mechanism of MDA-MB-231 cells using a kit and flow cytometer. As shown in fig. 5, the proportion of apoptotic and necrotic cells was not significantly affected by light alone or CTan alone treatment compared to the control group, further demonstrating that the light conditions employed in the present invention were not toxic to the cells. However, when light and CTan act synergistically, the proportion of apoptotic and necrotic cells, especially the proportion of apoptotic cells, increases by nearly 20-fold compared to the other groups, so that the results of this section of experiment indicate that the CTan photodynamic effect mediated anti-MDA-MB-231 cell effect is achieved by effectively inducing apoptosis and necrosis.
Example 6
Effects of CTan photosensitization on intracellular ROS levels in MDA-MB-231 cells:
after MDA-MB-231 cells were incubated with 2. Mu.g/mLCTan for 4 hours, they were then irradiated with a 30WLED lamp plate having a center wavelength of 460nm for 30 minutes, and the cells were treated after the irradiation was completed and analyzed by flow cytometry. Wherein, (A) is a control group, (B) is an independent illumination group, (C) is an independent drug group, and (D) is the combined action of the drugs and illumination, and the specific result is shown in FIG. 6.
In particular MDA-MB-231 cell suspension at 1X 10 5 Density of wells/plates in 6-well plates, cells were placed in 5% CO2 in an incubator at 37℃for 24h, then incubated with 2. Mu.g/mLCTan for 4h, then irradiated with 30WLED lamp plates with a center wavelength of 460nm for 30min, the supernatant removed, washed twice with PBS, 0.25% pancreatin solution (without EDTA) 0.7mL was added per well, the cells were collected by digestion, resuspended in 1mL serum-free medium containing DCF-DA 10. Mu. Mol/L, incubated in an incubator at 37℃for 20min with 1 inversion every 3min, then washed 3 times with serum-free medium, and analyzed by flow cytometry after 200. Mu. LPBS resuspension.
As can be seen from fig. 6: the invention has been demonstrated in the foregoing in O 2 In the presence of CTan, is capable of generating 1 O 2 It is demonstrated that CTan is capable of producing a photodynamic effect upon absorption of light. At the same time, CTan can cross cell membrane to enter cell interior, and the photochemical reaction of CTan in cell can affect the oxidation pressure change in MDA-MB-231 cell interior, so that the invention next examines the photosensitization of CTan to MDA-MB-231 cell interiorEffects of ROS levels. The invention adopts DCFH-DA as the ROS probe and uses a flow cytometer for analysis. As shown in fig. 6, the fluorescence intensity of DCF was close to that of the light alone group, which suggests that light alone did not cause changes in MDA-MB-231 intracellular ROS, and that drug release of CTan alone was able to increase MDA-MB-231 intracellular ROS levels compared to the control group, which may be related to the anticancer activity of CTan itself. Studies have been reported to demonstrate that CTan can increase intracellular ROS levels in a variety of cancer cells, thereby inhibiting the growth of cancer cells. Light and CTan synergy can further increase the intracellular ROS levels in MDA-MB-231 cells compared to CTan alone. The results of ROS level analysis indicate that the CTan photodynamic effect can increase intracellular ROS levels in MDA-MB-231 cells.
Example 7
CTan photosensitization mediated changes in mitochondrial membrane potential in MDA-MB-231 cells: wherein A is a control group, B is a single light group, C is a single cryptotanshinone group (2 mug/mL), and D is a CTan and light synergistic effect group (CTan concentration 2 mug/mL). MDA-MB-231 cells are incubated for 4 hours after CTan treatment of 2 mug/mL, then are irradiated for 30 minutes by a 30WLED lamp plate with the center wavelength of 460nm, and are dyed according to the procedure of the mitochondrial membrane potential detection kit instruction book, and then are analyzed by a flow cytometer.
As can be seen from fig. 7: the decrease of mitochondrial membrane potential is early marker time of apoptosis, and in order to further study the mechanism of CTan photosensitization and anticancer, the invention adopts JC-1 probe to detect the change of mitochondrial membrane potential of MDA-MB-231 cells, and when the mitochondrial membrane potential is decreased, the fluorescence of JC-1 probe is changed from red to green. The invention examines the change situation of the mitochondrial membrane potential of MDA-MB-231 cells after the illumination is finished and the culture is continued for 24 hours. As shown, the cell mitochondrial membrane potential of the non-illuminated group was similar, while the cell mitochondrial membrane potential of the illuminated group alone was similar to that of the control group, which indicated that at the drug concentrations employed in this experiment, the drug alone acted for 24 hours without causing a change in the cell mitochondrial membrane potential, while the irradiation light source employed herein did not affect the cell mitochondrial membrane potential. Then when light and CTan are cooperated, the mitochondrial membrane potential of MDA-MB-231 cells is changed obviously, namely, CTan photosensitization reduces the mitochondrial membrane potential of MDA-MB-231 cells obviously.
Example 8
Breast cancer MDA-MB-231 cells were incubated with 2. Mu.g/mLCtan for 4 hours, then irradiated with 30WLED lamp plate with a center wavelength of 460nm for 30min, and the cells were treated after the irradiation was completed for 24 hours for further culture, and analyzed by flow cytometry.
As can be seen from fig. 8: to further investigate the mechanism by which CTan photodynamic is resistant to MDA-MB-231 cells, the present invention analyzed the effect of CTan photosensitization on MDA-MB-231 cell cycle distribution. As shown, the light treatment alone and the CTan treatment alone hardly affected the periodic distribution of MDA-MB-231 cells compared to the control group, and the light irradiation was performed after the CTan was incubated with the cells for 4 hours, significantly reducing the proportion of cells in the G1 phase, while the proportion of cells in the G2 phase was greatly increased. These results indicate that the photodynamic effect of CTan causes G2/M arrest in MDA-MB-231 cells.
Claims (3)
1. Use of cryptotanshinone in the preparation of a photodynamic therapeutic agent, characterized in that: the cryptotanshinone acts as a photosensitizer activated with long wavelength light between 460 and 520 nm.
2. The application of cryptotanshinone in preparing a medicine for treating triple negative breast cancer by photodynamic therapy is characterized in that: the cryptotanshinone acts as a photosensitizer activated with long wavelength light between 460 and 520 nm.
3. Use according to claim 1 or 2, characterized in that: after the photo-activation of cryptotanshinone, free radical oxygen species with cytotoxicity are released.
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| 刘伟 等."丹参酮胶囊联合氨基酮戊酸光动力疗法对痤疮患 者血清IL-8、TNF-α 和皮脂分泌的影响".《中国中西医结合皮肤性病学杂志》.2018,第17卷(第5期),第426-428页. * |
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