CN109875994A - II type polyketide of halogenation is inhibiting the application in hepatoma cell proliferation - Google Patents
II type polyketide of halogenation is inhibiting the application in hepatoma cell proliferation Download PDFInfo
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- CN109875994A CN109875994A CN201910290460.5A CN201910290460A CN109875994A CN 109875994 A CN109875994 A CN 109875994A CN 201910290460 A CN201910290460 A CN 201910290460A CN 109875994 A CN109875994 A CN 109875994A
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Abstract
The present invention provides II type polyketides of halogenation to inhibit the application in hepatoma cell proliferation, integrin regulates and controls liver cancer growth, invasion and transfer, it is the important target spot of hepatoma-targeting drug therapy, II type polyketide of halogenation passes through in conjunction with liver cancer cells surface receptor integrin a subunit LFA-1a/ITGAL, exciting integrin LFA-1, it improves PI3K/Akt access regulation downstream hepatoma cell apoptosis and divides relevant gene expression dose, to inhibit hepatoma cell proliferation.
Description
Technical field
The invention belongs to genetically engineered drug fields, and in particular to II type polyketide of halogenation is inhibiting liver cancer cells
Application in proliferation.
Background technique
Liver cancer seriously threatens human life and health, and exploring treatment liver cancer new method is global problem.Primary carcinoma of liver
(Primary Hepatic Carcinoma, PHC) is one of malignant liver common in the world.Newly suffer from PHC people in the annual whole world
Number occupies global malignant liver disease incidence the 6th, the cause of death the 3rd because PHC died is up to 59.8 ten thousand people for 62.6 ten thousand people
Position.And it newly sends out in liver cancer case 55% and betides China, and liver cancer case fatality rate occupies the 2nd in various cancer case fatality rate.Base
It constitutes a serious threat in liver cancer to national health, explores more efficiently treatment method and be one and urgently to be resolved worldwide ask
Topic.Application No. is 201610394737.5,106167495 A of Publication No. CN, a kind of entitled halogenation II type polyketone class antibiosis
Plain, preparation method and applications patents disclose a kind of II type polyketide of halogenation, and resisting liver cancer activity test shows this
Good resisting liver cancer activity is all shown to the cell strain of liver cancer separate sources, provides very high research valence for clinical research
Value.
Summary of the invention
The object of the present invention is to provide II type polyketides of halogenation to inhibit the application in hepatoma cell proliferation, to suppression
Hepatoma cell proliferation processed has remarkable result.
The technical scheme adopted by the invention is that II type polyketide of halogenation is inhibiting answering in hepatoma cell proliferation
With.
Other of the invention are further characterized in that,
Inhibiting the application in HepG2 hepatoma cell proliferation more particularly to II type polyketide of halogenation.
Inhibiting the application in Hep1 hepatoma cell proliferation more particularly to II type polyketide of halogenation.
Inhibiting the application in MHCC97H hepatoma cell proliferation more particularly to II type polyketide of halogenation.
The structure of II type polyketide of halogenation are as follows:
Beneficial effects of the present invention are, can by the way that II type polyketide of halogenation is used for clinical treatment liver cancer diseases
To inhibit hepatoma cell proliferation, very high researching value is provided to treat the clinical research of liver cancer.
Detailed description of the invention
Fig. 1 is a subunit LFA-1a/ITGAL of 1.0 μm of ol/L in conjunction with II type polyketide of various concentration halogenation
Fluorescent quenching effect analysis figure；
Fig. 2 is II type polyketide of halogenation to PPAR γ Gene Expression situation analysis figure；
Fig. 3 is II type polyketide of halogenation to MAPK Gene Expression situation analysis figure；
Fig. 4 is II type polyketide of halogenation to CREB Gene Expression situation analysis figure；
Fig. 5 is II type polyketide of halogenation to GSK3B Gene Expression situation analysis figure；
Fig. 6 is that cell number spirogram under preceding microscope is administered in HepG2 liver cancer cells；
Fig. 7 is cell number spirogram under microscope after the administration of HepG2 liver cancer cells；
Fig. 8 is that cell number spirogram under preceding microscope is administered in Hep1 liver cancer cells；
Fig. 9 is cell number spirogram under microscope after the administration of Hep1 liver cancer cells；
Figure 10 is that cell number spirogram under preceding microscope is administered in MHCC97H liver cancer cells；
Figure 11 is cell number spirogram under microscope after the administration of MHCC97H liver cancer cells；
Figure 12 is that cell number spirogram under preceding microscope is administered in L02 normal liver cell；
Figure 13 is cell number spirogram under microscope after the administration of L02 normal liver cell；
Figure 14 is influence diagram of the II type polyketide of halogenation to HepG2 liver cancer cells survival rate of various concentration；
Figure 15 is influence diagram of the II type polyketide of halogenation to Hep1 liver cancer cells survival rate of various concentration；
Figure 16 is influence diagram of the II type polyketide of halogenation to MHCC97H liver cancer cells survival rate of various concentration；
Figure 17 is influence diagram of the II type polyketide of halogenation to L02 normal liver cell survival rate of various concentration；
Specific embodiment
Analysis the present invention is based on II type polyketide of halogenation to hepatoma cell proliferation depression effect, below with reference to attached
The present invention is described in detail with specific embodiment for figure.
Integrin regulates and controls liver cancer growth, invasion and transfer, is the important target spot of hepatoma-targeting drug therapy, and II type of halogenation is poly-
Ketone antibiotic is by conjunction with liver cancer cells surface receptor integrin a subunit LFA-1a/ITGAL, exciting integrin LFA-1,
To make PI3K/Akt passage downstream promote the PPAR γ gene, MAPK gene, CREB gene of hepatoma cell apoptosis and division with
And GSK3The gene expression dose of 1 B gene improves, and accelerates apoptosis and the division of liver cancer cells, reaches and inhibits hepatoma cell proliferation
Effect.
As shown in Figure 1, the curve in figure successively represents a subunit LFA-1a/ITGAL difference of 1.0 μm of ol/L from top to bottom
With the fluorescent quenching of the II type polyketide of halogenation of 0,0.1,0.2,0.3,0.4,0.5,0.6,0.7,0.8,0.9 μm of ol/L
Effect analysis, it can be seen from the figure that fluorescence intensity is more and more weaker with the increase of II type polyketide concentration of halogenation,
Therefore, it is possible to judge that containing integrin a subunit LFA-1a/ITGAL's with the increase of II type polyketide concentration of halogenation
Protein is combined more and more, and remaining protein content is fewer and fewer, shows that II type polyketide of halogenation and a are sub-
There is interaction between base LFA-1a/ITGAL.
As shown in Fig. 2 to 5, Control represents the cell controls that II type polyketide of halogenation is not added；8h,12h,16h
Represent the incubation of II type polyketide of halogenation 8h, 12h, 16h that concentration is added as 20 μM/ml to cell.
PPAR γ gene, MAPK gene, CREB gene and GSK31 B gene is the downstream gene of PI3K/Akt access,
And above four kinds of genes are the rush apoptogenes of liver cancer cells, it can be seen from the figure that II type polyketone class antibiosis of halogenation is added
After element, PAR γ gene, MAPK gene, CREB gene and GSK3The gene expression dose of B is significantly improved, so as to
To judge that II type polyketide of halogenation can promote PPAR γ gene, MAPK gene, CREB gene and GSK31 B gene
Gene expression dose
Essence in order to better understand the present invention, below will be with for the pharmacological evaluation of II type polyketide of halogenation
Bright its is inhibiting the application in hepatoma cell proliferation.
(1) cell recovery: taking out -80 DEG C of human hepatoma cell strain HepG2, Hep1, MHCC97H frozen and normal hepatocytes are thin
Born of the same parents L02 is put into rapidly in 37 DEG C of water-baths and thaws；Cell suspension is transferred in the 15mL centrifuge tube of the complete medium containing 2mL,
800rpm is centrifuged 3min.It takes 1mL complete medium to suspend again the cell of precipitating, and is transferred to the thin of the complete medium containing 3mL
It is dispelled in born of the same parents' culture bottle uniformly, is placed in 37 DEG C of CO2Incubator culture.
(2) cell passes on: discarding the culture medium in culture bottle, PBS liquid cleans 2 times, and 1mL0.25% trypsase is added and disappears
Change and observe cellular morphology under inverted microscope, terminates digestion with the complete culture medium of 3mL when contraction is rounded.Gently dispel cell
Cell suspension is transferred to 15mL centrifuge tube by the attached cell in culture bottle, and 800rpm is centrifuged 3min.Take 1mL complete medium
Again the cell for the precipitating that suspends, and be transferred in the Tissue Culture Flask of the complete medium containing 3mL and dispel uniformly, it is placed in 37 DEG C, 5%
CO2It is cultivated in incubator.
(3) cell cryopreservation: taking out the cell for being spread to Tissue Culture Flask bottom area 80%, and PBS is cleaned 2 times, and 1mL is added
Cellular morphology is observed under 0.25% trypsin digestion inverted microscope, is terminated when contraction is rounded with the complete culture medium of 3mL
Digestion.The attached cell in Tissue Culture Flask is gently dispelled, cell suspension is transferred to 15mL centrifuge tube, 800rpm, centrifugation
3min.Cell precipitation is transferred in 1.8mL cryopreservation tube after being suspended again with 1mL frozen stock solution, 4 DEG C of placement 30min, -20 DEG C of placements
1h is placed in -80 DEG C of preservations.
(4) cell administration: taking out the cell of passage, discards the culture medium in culture bottle, and PBS is cleaned 2 times, and 1mL is added
0.25% trypsin digestion, when cellular contraction is rounded, the complete culture medium of 3mL terminates digestion.Cell is gently blown and beaten, is made
Attached cell in Tissue Culture Flask sufficiently falls off, and collects cell suspension and is transferred to 15mL centrifuge tube, 800rpm is centrifuged 3min.
It is 5 × 10 that cell precipitation complete medium, which adjusts cell concentration,4A/mL takes 100 μ L cell suspensions, is separately added into 96 orifice plates
Culture hole is placed in 37 DEG C of CO2Incubator culture is for 24 hours.When cell grows to the 80% of culture hole, take II type of halogenation of 6 μ L poly-
Ketone antibiotic mother liquor is added in 1.5mL centrifuge tube, is diluted to 1mL with complete medium.The drug hole is successively diluted to 2
μM, 4 μM, 8 μM, 16 μM, 32 μM, 64 μM, 128 μM, 8 concentration such as 256 μM as test concentrations, each medicine group is 6 parallel
Multiple holes, 37 DEG C, 5%CO2Incubator effect terminates to cultivate for 24 hours afterwards.
(5) absorbance measurement: discarding the liquid in 96 orifice plates, and PBS cleans each orifice plate to noresidue, it is complete that 90 μ L are added
Culture medium and 10 μ L cck8 reagents, 37 DEG C are continued to terminate culture after being incubated for 4h, and microplate reader measures OD value at 490nm.
The II type polyketide of halogenation of various concentration acts on, HepG2 liver cancer cells, Hep1 liver cancer cells,
MHCC97H liver cancer cells and L02 normal liver cell for 24 hours after, observe HepG2 liver cancer cells, Hep1 liver cancer cells, MHCC97H
The cell quantity variation of liver cancer cells and L02 normal liver cell for 24 hours, and use the shadow of cck8 measurement drug cell proliferation
It rings.
As shown in Fig. 6 to Figure 13, can be seen that from Fig. 6 to Figure 13 HepG2 liver cancer cells, Hep1 liver cancer cells and
After II type polyketide of halogenation is added in MHCC97H liver cancer cells, cell quantity is significantly reduced, and thin in L02 normal hepatocytes
After II type polyketide of halogenation is added in born of the same parents, cell quantity is only slightly reduced.
As shown in Figure 14 to Figure 17, from Figure 14 to Figure 17 in as can be seen that the concentration of II type polyketide of halogenation is got over
Greatly, the cell survival rate of HepG2 liver cancer cells, Hep1 liver cancer cells, MHCC97H liver cancer cells and L02 normal liver cell is just
It is lower, when the control of II type polyketide concentration of halogenation is between 2 μM/mL to 32 μM/mL, II type polyketone class antibiosis of halogenation
Element the cell survival rate of L02 normal liver cell is influenced it is smaller, and to HepG2 liver cancer cells, Hep1 liver cancer cells and
MHCC97H liver cancer cells survival rate is larger.
According to each group OD490The average and standard deviation of value, is calculated by formula inhibitory rate of cell growth.
IC50(half maximal inhibitory concentration) refers to the 503nhibiting concentration of drug.It can refer to
Show that a certain drug or substance (inhibitor) are inhibiting certain biological process, concentration or dosage when such as cell death half, IC50
Value can be used to measure the ability of drug-induced apoptosis, i.e. inducibility is stronger, and the numerical value is lower, also can be reversed and illustrate certain
Tolerance degree of the cell to drug.
Data Processing in Experiment uses SPSS18.0 and GraphPadPrism6.0 statistical software, calculates the half-suppressed dense of drug
Spend IC50, see Table 1 for details.Table 1 is II type polyketide of various concentration halogenation to the thin of different liver cancer cells and normal liver cell
Born of the same parents' inhibiting rate analytical table, from table 1 it follows that the concentration of II type polyketide of halogenation is higher, it is thin to HepG2 liver cancer
Born of the same parents, Hep1 liver cancer cells, the inhibiting rate of MHCC97H liver cancer cells are then higher.
Table 1
Claims (5)
1. II type polyketide of halogenation is inhibiting the application in hepatoma cell proliferation.
2. application as described in claim 1, which is characterized in that inhibiting more particularly to II type polyketide of halogenation
Application in HepG2 hepatoma cell proliferation.
3. application as described in claim 1, which is characterized in that inhibiting Hep1 more particularly to II type polyketide of halogenation
Application in hepatoma cell proliferation.
4. application as described in claim 1, which is characterized in that inhibiting more particularly to II type polyketide of halogenation
Application in MHCC97H hepatoma cell proliferation.
5. application as described in claim 1, which is characterized in that the structure of the II type polyketide of halogenation are as follows:
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114903887A (en) * | 2022-06-30 | 2022-08-16 | 延安大学 | Application of halogenated II type polyketone antibiotics in enhancing immunity |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434785A (en) * | 2016-06-08 | 2017-02-22 | 遵义医学院 | Preparation method of 2-n-heptyl-4-hydroxyquinoline |
| CN107937453A (en) * | 2017-11-22 | 2018-04-20 | 遵义医学院 | A kind of preparation method of II type halogenation polyketides of dichloro substitution and antibacterial activity application |
-
2019
- 2019-04-11 CN CN201910290460.5A patent/CN109875994A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434785A (en) * | 2016-06-08 | 2017-02-22 | 遵义医学院 | Preparation method of 2-n-heptyl-4-hydroxyquinoline |
| CN107937453A (en) * | 2017-11-22 | 2018-04-20 | 遵义医学院 | A kind of preparation method of II type halogenation polyketides of dichloro substitution and antibacterial activity application |
Non-Patent Citations (1)
| Title |
|---|
| 王荫荫: "卤化聚酮类抗生素zunyimycins抑菌及抗肿瘤活性初步分析", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114903887A (en) * | 2022-06-30 | 2022-08-16 | 延安大学 | Application of halogenated II type polyketone antibiotics in enhancing immunity |
| CN114903887B (en) * | 2022-06-30 | 2023-09-05 | 延安大学 | Application of halogenated type II polyketone antibiotics in enhancing immunity |
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Application publication date: 20190614 |