CN108707581A - A method of restoring Expressions of suppressor gene PTEN using DNA methylation enzyme inhibitor - Google Patents
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Abstract
The present invention relates to a kind of methods for restoring expression of tumor suppressor gene, are a kind of methods for restoring Expressions of suppressor gene PTEN using DNA methylation enzyme inhibitor specifically.DNA methylation enzyme inhibitor used in the present invention is 5 '-Aza, can restore the expression of mir-200 families using it, to promote the expression of PTEN to play the role of inhibiting tumor development.This method is based on the present invention newfound a kind of a kind of new mechanism of action and pattern of hostile miRNA, i.e. a kind of miRNA (miR-21a) can inhibit the expression of another miRNA (mir-200c), be suppressed so as to cause expression of tumor suppressor gene.
Description
Technical field
The present invention relates to a kind of methods for restoring expression of tumor suppressor gene, are a kind of to be inhibited using DNA methylation enzyme specifically
The method that agent restores Expressions of suppressor gene PTEN.
Background technology
Mir-200c family members have been proved to be able to adjust EMT in kinds of tumors.And previously there is research table
Bright, mir-200c can promote the expression of PTEN.And some researches show that the hyper-methylations on the islands CpG can mediate mir-200 recently
The regulation and control of the epigenetic of family, to adjust the transfer of EMT and tumour.We pass through MethPrimer-Design MSP/
MethPrimer software predictions on the websites BSP primers and predict CpG island mir-200c
The site (Fig 6C) on the possible islands CpG promoter, and methylate and the non-primer to methylate from 5 Dui provided, pass through
Methl-specific PCR (MSP) successfully have found a pair of of methylated primers and non-methylated primers carry out subsequent experimental.
And it can be seen from the figure that the RAW264.7 after normal RAW264.7 cells and primary macrophage and the domestication of 4T1 supernatants is thin
The methylation on the islands CpG is all high at the mir-200c promoter of born of the same parents or the gDNA of primary macrophage, and passes through 5-
After Aza processing, the degree that mir-200c promoter methylate is substantially reduced.
Mir-21a can trigger the silence of the mir-200c for the dependence that methylates.The mir- that the hyper-methylation on the islands CpG mediates
The silence of 200 family's epigenetics and the expression for lowering PTEN, promote M2 and transfer.We use methylation status of PTEN promoter
(MSP) it analyzes the promoter of mir-200, finds when mir-21a is overexpressed, the promoter of mir-200 is high methyl
Change.It is handled when with 5 '-Aza of DNA- demethylations reagent, it is thus opposite with the effect of mir-21a and mir-200 can be repaired
Transcription.This shows that mir-21a may play an important role during dynamic control DNA methylation.Mir-21a
Inhibition of the PTEN to PI3K/AKT signal paths of mimicl indirect down-regulations, in a disguised form has encouraged the growth of tumour cell.Change speech
It, it is believed that mir-21a is strictly to be played a role to adjust PI3K/Akt signal paths by adjusting PTEN.
Therefore, how to find that mir-21a inhibits the mechanism of PTEN expression, and utilize this mechanism, realize and restore PTEN tables
It reaches, to inhibit the occurrence and development of cancer that there is important value.
Invention content
The purpose of the present invention：
The present invention is directed to explore a kind of method for restoring Expressions of suppressor gene PTEN using DNA methylation enzyme inhibitor.
The technical principle of the present invention：
On the one hand Mir-21a can be directly targeted the 3 ' UTR regions of PTEN, to lower the expression of PTEN, and then influence
PI3K/AKT signal paths, to influence the polar conversion of macrophage, to be more favorable for the migration and increasing of tumour cell
It grows；On the other hand, mir-21a can also trigger the islands the CpG hyper-methylation of mir-200c promoter, to inhibit mir-
The expression of 200c, and then the expression of PTEN is inhibited indirectly.According to above-mentioned principle, we use DNA methylation enzyme inhibitor 5 '-
Aza restores the expression of mir-200 families, and then restores the expression of PTEN, to realize the purpose of control tumor development.
The present invention is achieved by the following technical solutions：
Using the 5 '-Aza normal RAW264.7 cells handled and primary macrophage, compare the mir- of the cell after processing
The expression of the methylation and Anti-oncogene PTEN on the islands CpG at 200c promoter, assessment use DNA methylation enzyme
The method that 5 '-Aza of inhibitor restores Expressions of suppressor gene PTEN.
Description of the drawings
Mir-200c's opens in Fig. 1 Methl-specific PCR detection RAW264.7 cell lines and primary macrophage
Mover methylation status.
It can be seen that at normal RAW264.7 cells and primary macrophage mir-200c promoter the islands CpG the journey that methylates
Degree is all high, and after 5-Aza is handled, the degree that mir-200c promoter methylate is substantially reduced.
Fig. 2 determines 5 '-AZa activities of DNA methylation enzyme inhibitor by cytotoxicity.
It can be seen that under the concentration of 2 every liter of micromoles, cell can keep 60% activity, in line with improving 5 '-AZa as far as possible
Activity and save the principle of reagent, determine that every liter of 2 micromole is used as activity.
Fig. 3 determines the action time of 5 '-AZa of DNA methylation enzyme inhibitor by real time PCR methods
Using 2 moles every liter of 5 '-Aza function cells, handle the different time (i.e. 0h, for 24 hours, 48h, 72h, 84h and
96h), detection detects the expression of pri-200c using the method for real time PCR.It can be seen that having obtained highest table in 84h
Up to amount.Therefore the 5 '-Aza of 2 every liter of micromoles is selected to act on action times of the 84h as DNA methylation enzyme inhibitor.
Fig. 4 restores Expressions of suppressor gene PTEN using 5 '-Aza in Real time PCR assessment RAW264.7 cell lines
As a result.
Real time PCR's the result shows that, with 5-Aza handle through 4T1 supernatants domestication after RAW264.7 in mir-21a
Expression do not change substantially, and the expression of mir-200c, mir-200a, PTEN are obviously raised.
Fig. 5 restores the knot of Expressions of suppressor gene PTEN using 5 '-Aza in Real time PCR assessment primary macrophages
Fruit.
Real time PCR's the result shows that, with 5-Aza handle through 4T1 supernatants domestication after primary macrophage in
The expression of mir-21a does not change substantially, and the expression of mir-200c, mir-200a, PTEN are obviously raised.
Specific implementation mode
One, determine that methylation changes
Research shows that the hyper-methylation on the islands CpG can mediate the regulation and control of the epigenetic of mir-200 families, to adjust
The transfer of EMT and tumour.We pass through MethPrimer-Design MSP/BSP primers and in the present invention
MethPrimer software predictions on the websites the predict CpG island possible islands CpG mir-200c promoter
Site, and methylate and the non-primer to methylate from 5 Dui provided, it is successfully looked for by methl-specific PCR (MSP)
It has arrived a pair of of methylated primers and non-methylated primers carries out subsequent experimental.It is shown from the result of study, normal RAW264.7 is thin
The methylation on the islands CpG is all high at born of the same parents and primary macrophage mir-200c promoter, and after 5-Aza is handled,
The degree that mir-200c promoter methylate is substantially reduced.(as shown in Figure 1)
Two, the activity of 5 '-AZa of DNA methylation enzyme inhibitor is determined
The present invention determines the activity of 5 '-AZa of DNA methylation enzyme inhibitor by cellulotoxic experiment, the cell used
System is CCK8.0.1 every liter of micromole, 0.5 every liter of micromole, 1 every liter of micromole, 2 every liter of micromoles, 4 micromoles are chosen respectively
Every liter, every liter of 8 every liter of micromoles and 16 micromoles are compared assessment cytotoxicity.Obtain that the results are shown in Figure 2, in line with to the greatest extent
The activity of 5 '-AZa may be improved and save the principle of reagent, we have selected every liter of 2 micromole as activity.
Three, the action time of 5 '-AZa of DNA methylation enzyme inhibitor is determined
The present invention assesses the action time of 5 '-AZa of DNA methylation enzyme inhibitor by the expression quantity of pri-200c.Specifically
Method is as follows, using the 5 '-Aza function cells of 2 every liter of micromoles, handle the different time (i.e. 0h, for 24 hours, 48h, 72h, 84h
And 96h), detection detects the expression of pri-200c using the method for real time PCR.Obtain that the results are shown in Figure 3, it can be with
It determines in 84h, the expression quantity of pri-200c is maximum.Therefore the 5 '-Aza of 2 every liter of micromoles is selected to act on 84h as DNA first
The action time of base enzyme inhibitor.
Embodiment 1
Using present invention determine that 5 '-Aza of DNA methylation enzyme inhibitor 2 every liter of micromole of activity and effect when
Between 84 hours, processing by domestication RAW264.7 cell line samples 1.After treatment extracts the total serum IgE of cell line sample,
Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then expands mir-21a, mir- respectively using the primer of specificity
Six genes such as 200c, PTEN, p53, CCL2, VEGFa are recorded using Ct values as the foundation of expression quantity.
The 5 '-Aza processing of DNA methylation enzyme inhibitor is not passed through using same method extraction control group simultaneously
The total serum IgE of RAW264.7 cell line samples 1.Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, is then used special
Property primer expand mir-21a respectively, mir-200c, PTEN, p53, CCL2, six genes such as VEGFa, using Ct values as expression
The foundation of amount, is also recorded.Then the expression quantity of experimental group is compared to obtain experimental group with the expression quantity of control group
Relative expression quantity, the relative expression quantity be control group expression quantity as 1 when relative value.
Obtaining result is：
| Gene | mir-21a | mir-200c | PTEN | p53 | CCL2 | VEGFa |
| Expression quantity | 1.18 | 1.80 | 1.77 | 3.82 | 0.31 | 0.30 |
As a result show that mir-21a in RAW264.7 cell line samples 1, mir-200c, PTEN, p53 high are expressed, CCL2,
VEGFa low expressions.
Embodiment 2
Using present invention determine that 5 '-Aza of DNA methylation enzyme inhibitor 2 every liter of micromole of activity and effect when
Between 84 hours, processing by domestication RAW264.7 cell line samples 2.After treatment extracts the total serum IgE of cell line sample,
Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then expands mir-21a, mir- respectively using the primer of specificity
Six genes such as 200c, PTEN, p53, CCL2, VEGFa are recorded using Ct values as the foundation of expression quantity.
The 5 '-Aza processing of DNA methylation enzyme inhibitor is not passed through using same method extraction control group simultaneously
The total serum IgE of RAW264.7 cell line samples 2.Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, is then used special
Property primer expand mir-21a respectively, mir-200c, PTEN, p53, CCL2, six genes such as VEGFa, using Ct values as expression
The foundation of amount, is also recorded.Then the expression quantity of experimental group is compared to obtain experimental group with the expression quantity of control group
Relative expression quantity, the relative expression quantity be control group expression quantity as 1 when relative value.
Obtaining result is：
| Gene | mir-21a | mir-200c | PTEN | p53 | CCL2 | VEGFa |
| Expression quantity | 1.19 | 1.83 | 1.79 | 3.80 | 0.33 | 0.31 |
As a result show that mir-21a in RAW264.7 cell line samples 2, mir-200c, PTEN, p53 high are expressed, CCL2,
VEGFa low expressions.
Embodiment 3
Using present invention determine that 5 '-Aza of DNA methylation enzyme inhibitor 2 every liter of micromole of activity and effect when
Between 84 hours, processing by domestication RAW264.7 cell line samples 3.After treatment extracts the total serum IgE of cell line sample,
Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then expands mir-21a, mir- respectively using the primer of specificity
Six genes such as 200c, PTEN, p53, CCL2, VEGFa are recorded using Ct values as the foundation of expression quantity.
The 5 '-Aza processing of DNA methylation enzyme inhibitor is not passed through using same method extraction control group simultaneously
The total serum IgE of RAW264.7 cell line samples 3.Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, is then used special
Property primer expand mir-21a respectively, mir-200c, PTEN, p53, CCL2, six genes such as VEGFa, using Ct values as expression
The foundation of amount, is also recorded.Then the expression quantity of experimental group is compared to obtain experimental group with the expression quantity of control group
Relative expression quantity, the relative expression quantity be control group expression quantity as 1 when relative value.
Obtaining result is：
| Gene | mir-21a | mir-200c | PTEN | p53 | CCL2 | VEGFa |
| Expression quantity | 1.19 | 1.80 | 1.80 | 3.85 | 0.33 | 0.31 |
As a result show that mir-21a in RAW264.7 cell line samples 3, mir-200c, PTEN, p53 high are expressed, CCL2,
VEGFa low expressions.
Embodiment 4
Using present invention determine that 5 '-Aza of DNA methylation enzyme inhibitor 2 every liter of micromole of activity and effect when
Between 84 hours, processing by domestication RAW264.7 cell line samples 4.After treatment extracts the total serum IgE of cell line sample,
Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then expands mir-21a, mir- respectively using the primer of specificity
Six genes such as 200c, PTEN, p53, CCL2, VEGFa are recorded using Ct values as the foundation of expression quantity.
The 5 '-Aza processing of DNA methylation enzyme inhibitor is not passed through using same method extraction control group simultaneously
The total serum IgE of RAW264.7 cell line samples 4.Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, is then used special
Property primer expand mir-21a respectively, mir-200c, PTEN, p53, CCL2, six genes such as VEGFa, using Ct values as expression
The foundation of amount, is also recorded.Then the expression quantity of experimental group is compared to obtain experimental group with thin expression quantity is compareed
Relative expression quantity, the relative expression quantity be control group expression quantity as 1 when relative value.
Obtaining result is：
| Gene | mir-21a | mir-200c | PTEN | p53 | CCL2 | VEGFa |
| Expression quantity | 1.19 | 1.85 | 1.86 | 3.80 | 0.34 | 0.33 |
As a result show that mir-21a in RAW264.7 cell line samples 4, mir-200c, PTEN, p53 high are expressed, CCL2,
VEGFa low expressions.
Embodiment 5
Using present invention determine that 5 '-Aza of DNA methylation enzyme inhibitor 2 every liter of micromole of activity and effect when
Between 84 hours, processing by domestication RAW264.7 cell line samples 5.After treatment extracts the total serum IgE of cell line sample,
Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then expands mir-21a, mir- respectively using the primer of specificity
Six genes such as 200c, PTEN, p53, CCL2, VEGFa are recorded using Ct values as the foundation of expression quantity.
The 5 '-Aza processing of DNA methylation enzyme inhibitor is not passed through using same method extraction control group simultaneously
The total serum IgE of RAW264.7 cell line samples 5.Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, is then used special
Property primer expand mir-21a respectively, mir-200c, PTEN, p53, CCL2, six genes such as VEGFa, using Ct values as expression
The foundation of amount, is also recorded.Thin expression quantity will be then tested to be compared to obtain experimental group with the expression quantity of control group
Relative expression quantity, the relative expression quantity be control group expression quantity as 1 when relative value.
Obtaining result is：
| Gene | mir-21a | mir-200c | PTEN | p53 | CCL2 | VEGFa |
| Expression quantity | 1.19 | 1.86 | 1.82 | 3.80 | 0.33 | 0.30 |
As a result show that mir-21a in RAW264.7 cell line samples 5, mir-200c, PTEN, p53 high are expressed, CCL2,
VEGFa low expressions.
Embodiment 6
Using present invention determine that 5 '-Aza of DNA methylation enzyme inhibitor 2 every liter of micromole of activity and effect when
Between 84 hours, processing by domestication primary macrophage system sample 1.After treatment extracts the total serum IgE of cell line sample,
Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then expands mir-21a, mir- respectively using the primer of specificity
Six genes such as 200c, PTEN, p53, CCL2, VEGFa are recorded using Ct values as the foundation of expression quantity.
The primary of DNA methylation enzyme inhibitor 5 '-Aza processing is not passed through using same method extraction control group simultaneously
The total serum IgE of macrophage system sample 1.Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then uses specificity
Primer expands mir-21a, mir-200c, PTEN, p53, CCL2, six genes such as VEGFa, using Ct values as expression quantity respectively
Foundation is also recorded.Then the expression quantity of experimental group is compared to obtain the opposite of experimental group with the expression quantity of control group
Expression quantity, the relative expression quantity be control group expression quantity as 1 when relative value.
Obtaining result is：
| Gene | mir-21a | mir-200c | PTEN | p53 | CCL2 | VEGFa |
| Expression quantity | 1.19 | 1.65 | 1.78 | 1.78 | 0.21 | 1.00 |
As a result show that mir-21a, mir-200c in primary macrophage system sample 1, PTEN, p53 high are expressed, CCL2,
VEGFa low expressions.
Embodiment 7
Using present invention determine that 5 '-Aza of DNA methylation enzyme inhibitor 2 every liter of micromole of activity and effect when
Between 84 hours, processing by domestication primary macrophage system sample 2.After treatment extracts the total serum IgE of cell line sample,
Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then expands mir-21a, mir- respectively using the primer of specificity
Six genes such as 200c, PTEN, p53, CCL2, VEGFa are recorded using Ct values as the foundation of expression quantity.
The primary of DNA methylation enzyme inhibitor 5 '-Aza processing is not passed through using same method extraction control group simultaneously
The total serum IgE of macrophage system sample 2.Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then uses specificity
Primer expands mir-21a, mir-200c, PTEN, p53, CCL2, six genes such as VEGFa, using Ct values as expression quantity respectively
Foundation is also recorded.Then the expression quantity of experimental group is compared to obtain the opposite of experimental group with the expression quantity of control group
Expression quantity, the relative expression quantity be control group expression quantity as 1 when relative value.
Obtaining result is：
| Gene | mir-21a | mir-200c | PTEN | p53 | CCL2 | VEGFa |
| Expression quantity | 1.22 | 1.68 | 1.81 | 1.81 | 0.22 | 1.01 |
As a result show that mir-21a, mir-200c in primary macrophage system sample 2, PTEN, p53 high are expressed, CCL2,
VEGFa low expressions.
Embodiment 8
Using present invention determine that 5 '-Aza of DNA methylation enzyme inhibitor 2 every liter of micromole of activity and effect when
Between 84 hours, processing by domestication primary macrophage system sample 3.After treatment extracts the total serum IgE of cell line sample,
Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then expands mir-21a, mir- respectively using the primer of specificity
Six genes such as 200c, PTEN, p53, CCL2, VEGFa are recorded using Ct values as the foundation of expression quantity.
The primary of DNA methylation enzyme inhibitor 5 '-Aza processing is not passed through using same method extraction control group simultaneously
The total serum IgE of macrophage system sample 3.Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then uses specificity
Primer expands mir-21a, mir-200c, PTEN, p53, CCL2, six genes such as VEGFa, using Ct values as expression quantity respectively
Foundation is also recorded.Then the expression quantity of experimental group is compared to obtain the opposite of experimental group with the expression quantity of control group
Expression quantity, the relative expression quantity be control group expression quantity as 1 when relative value.
Obtaining result is：
| Gene | mir-21a | mir-200c | PTEN | p53 | CCL2 | VEGFa |
| Expression quantity | 1.23 | 1.68 | 1.85 | 1.80 | 0.20 | 1.03 |
As a result show that mir-21a, mir-200c in primary macrophage system sample 3, PTEN, p53 high are expressed, CCL2,
VEGFa low expressions.
Embodiment 9
Using present invention determine that 5 '-Aza of DNA methylation enzyme inhibitor 2 every liter of micromole of activity and effect when
Between 84 hours, processing by domestication primary macrophage system sample 4.After treatment extracts the total serum IgE of cell line sample,
Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then expands mir-21a, mir- respectively using the primer of specificity
Six genes such as 200c, PTEN, p53, CCL2, VEGFa are recorded using Ct values as the foundation of expression quantity.
The primary of DNA methylation enzyme inhibitor 5 '-Aza processing is not passed through using same method extraction control group simultaneously
The total serum IgE of macrophage system sample 4.Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then uses specificity
Primer expands mir-21a, mir-200c, PTEN, p53, CCL2, six genes such as VEGFa, using Ct values as expression quantity respectively
Foundation is also recorded.Then the expression quantity of experimental group is compared to obtain the opposite of experimental group with the expression quantity of control group
Expression quantity, the relative expression quantity be control group expression quantity as 1 when relative value.
Obtaining result is：
| Gene | mir-21a | mir-200c | PTEN | p53 | CCL2 | VEGFa |
| Expression quantity | 1.21 | 1.71 | 1.83 | 1.83 | 0.20 | 1.05 |
As a result show that mir-21a, mir-200c in primary macrophage system sample 4, PTEN, p53 high are expressed, CCL2,
VEGFa low expressions.
Embodiment 10
Using present invention determine that 5 '-Aza of DNA methylation enzyme inhibitor 2 every liter of micromole of activity and effect when
Between 84 hours, processing by domestication primary macrophage system sample 5.After treatment extracts the total serum IgE of cell line sample,
Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then expands mir-21a, mir- respectively using the primer of specificity
Six genes such as 200c, PTEN, p53, CCL2, VEGFa are recorded using Ct values as the foundation of expression quantity.
The primary of DNA methylation enzyme inhibitor 5 '-Aza processing is not passed through using same method extraction control group simultaneously
The total serum IgE of macrophage system sample 5.Reverse transcription is carried out using Reverse Transcriptase kit and obtains total cDNA, then uses specificity
Primer expands mir-21a, mir-200c, PTEN, p53, CCL2, six genes such as VEGFa, using Ct values as expression quantity respectively
Foundation is also recorded.Then the expression quantity of experimental group is compared to obtain the opposite of experimental group with the expression quantity of control group
Expression quantity, the relative expression quantity be control group expression quantity as 1 when relative value.
Obtaining result is：
| Gene | mir-21a | mir-200c | PTEN | p53 | CCL2 | VEGFa |
| Expression quantity | 1.21 | 1.68 | 1.80 | 1.85 | 0.23 | 1.05 |
As a result show that mir-21a, mir-200c in primary macrophage system sample 5, PTEN, p53 high are expressed, CCL2,
VEGFa low expressions.
Summarize embodiment 1-5 to obtain restoring suppression cancer using 5 '-Aza in Real time PCR assessment RAW264.7 cell lines
PTEN Gene expression as a result, as shown in Figure 4.It can be seen that handling mir- in the RAW264.7 after the domestication of 4T1 supernatants with 5-Aza
The expression of 21a does not change substantially, and the expression of mir-200c, mir-200a, PTEN are obviously raised.
Summarize embodiment 6-10 to obtain restoring suppression cancer base using 5 '-Aza in Real time PCR assessment primary macrophages
Because PTEN expression is as a result, as shown in Figure 5.It can be seen that handling mir- in the primary macrophage after the domestication of 4T1 supernatants with 5-Aza
The expression of 21a does not change substantially, and the expression of mir-200c, mir-200a, PTEN are obviously raised.
Claims (5)
1. a kind of method for restoring Expressions of suppressor gene PTEN using DNA methylation enzyme inhibitor, which is characterized in that be a kind of logical
It crosses DNA methylation enzyme inhibitor and restores a kind of expression of correlation function miRNA, restore the method for expression of tumor suppressor gene.
2. a kind of method for restoring Expressions of suppressor gene PTEN using DNA methylation enzyme inhibitor according to claim 1,
It is characterized in that, the DNA methylation enzyme inhibitor used is 5 '-Aza.
3. a kind of method for restoring Expressions of suppressor gene PTEN using DNA methylation enzyme inhibitor according to claim 1,
It is characterized in that, restoring a kind of expression of correlation function miRNA, to restore expression of tumor suppressor gene, the miRNA restored is
mir-200c。
4. a kind of method for restoring Expressions of suppressor gene PTEN using DNA methylation enzyme inhibitor according to claim 1,
It is characterized in that, 2 every microlitre of the micromole of activity of 5 '-Aza, action time is 84 hours.
5. a kind of method for restoring Expressions of suppressor gene PTEN using DNA methylation enzyme inhibitor according to claim 1,
It is characterized in that the action site of DNA methylation enzyme inhibitor is islands CpG at 5 '-Azamir-200c promoter on chromosome
DNA fragmentation.
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| CN112877309A (en) * | 2021-02-19 | 2021-06-01 | 北京大学 | N-terminal extended PTEN subtype PTEN zeta protein and coding gene and application thereof |
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| CN112877309A (en) * | 2021-02-19 | 2021-06-01 | 北京大学 | N-terminal extended PTEN subtype PTEN zeta protein and coding gene and application thereof |
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