CN105779452A - Oligonucleotide capable of inhibiting tumor growth and application of oligonucleotide - Google Patents

Abstract

The invention discloses an oligomeric nucleic acid for inhibiting tumor growth and its application. The oligonucleotide provided by the present invention is as follows (A) or (B): (A) double-stranded RNA: the double-stranded RNA is made up of sense strand (sequence 1) and antisense strand (sequence 2); (B) A chimeric duplex formed by complementary pairing of a single-stranded RNA (sequence 1) and a single-stranded DNA (sequence shown after replacing U in sequence 2 with T). The oligonucleotide provided by the invention can effectively inhibit tumor growth in vivo at the animal level; at the cell level, it can inhibit tumor cell proliferation, tumor cell invasion, tumor cell migration and tumor cell apoptosis. Therefore, the oligonucleotide provided by the present invention, as a novel small nucleic acid antitumor drug, can not only inhibit the growth of tumors in the body, reduce the risk of tumor spread, but also is expected to be used to prevent residual tumors after tumor resection or radiotherapy and chemotherapy. cells that cause tumor recurrence.

Description

A kind of oligonucleotide for inhibiting tumor growth and its application

technical field

The invention belongs to the field of biomedicine, and relates to an oligomeric nucleic acid for inhibiting tumor growth and its application.

Background technique

MicroRNA is a class of non-coding single-stranded RNA molecules encoded by endogenous genes with a length of about 19-25 nucleotides. It has high homology among different species and may regulate 1/3 of human genes. microRNAs participate in the regulation of individual development, and there are three main ways of cell proliferation, differentiation, cycle and other activities: mature microRNA complements and binds to mRNA, RISC specifically degrades target mRNA and inhibits mRNA translation; mature microRNA binds to target mRNA without affecting mRNA stability The 3'UTR of the target mRNA inhibits the expression of the target mRNA; after the microRNA cleaves the polyadenylate end of the target mRNA, the target mRNA is hydrolyzed by the 3' exonuclease, and the target mRNA is negatively regulated at the post-transcriptional level. By targeting different genes, microRNA can regulate the function of tumor cells. Studies have shown that more and more proto-oncogenes or tumor suppressor genes have significant differences in expression between tumor cells and normal tissues, and are associated with the occurrence and development of tumors. Using the function of proto-oncogene or tumor-suppressor gene of microRNA and related technologies may realize the possibility of tumor-targeted therapy.

Contents of the invention

The first object of the present invention is to provide an oligonucleotide.

The oligonucleotide provided by the present invention is as follows (A) or (B):

(A) double-stranded RNA: the double-stranded RNA is composed of a sense strand and an antisense strand, the sense strand has the sequence shown in sequence 1 in the sequence listing, and the antisense strand has the sequence shown in sequence 2 in the sequence listing;

(B) A chimeric duplex formed by complementary pairing of a single-stranded RNA and a single-stranded DNA: the single-stranded RNA in the chimeric duplex has the sequence shown in Sequence 1 in the sequence table, and the single-stranded DNA It has the sequence shown after replacing U in sequence 2 in the sequence listing with T.

In the present invention, in (A), the sequence of the sense strand is specifically sequence 1 in the sequence listing, and the sequence of the antisense strand is specifically sequence 2 in the sequence listing. In (B), the sequence of the single-stranded RNA is specifically sequence 1 in the sequence listing, and the sequence of the single-stranded DNA is specifically the sequence shown after replacing U in sequence 2 in the sequence listing with T.

The oligonucleotide can be modified as required by at least one of the following:

(1) Modification of the phosphodiester bond connecting the nucleotides in the nucleotide sequence of the oligonucleotide (such as the oxygen in the phosphodiester bond is replaced by sulfur);

(2) Modification of the 2'-OH of the ribose in the nucleotide sequence of the oligonucleotide (such as 2'-OH is substituted by methoxy, fluorine or deoxygenated);

(3) Modifications to the bases in the nucleotide sequence of the oligonucleotide (such as modifications such as LNA; modifications such as cholesterol, methoxy, thio, amino, etc.; 3' or 5' special modifications, including biological pigments, fluorophores, and other special labels).

Further, the oligomeric nucleic acid is chemically synthesized, and can carry out 2' fluoro (2'-F), 2' methoxy (2'-OMe), thio (PS) and 2' deoxy (2'-deoxy ) chemical modification. The modified oligonucleotide molecules can be respectively represented as follows: the oligonucleotide-2'-F, the oligonucleotide-PS, the oligonucleotide-2'-OMe and the oligonucleotide-2'- deoxy. Or 5'p, 5'thiol, 5'NH ₂ , 5'Chol, 5-Me-dC modification, 5'TET, 5'Biotin3'NH2, 3'Chol, 3'BHQ-1 of the nucleotide sequence , 3'Dabcyl, or C6S-S modification and other modifications. The oligomeric nucleic acid molecule is effective in important tumor regulatory genes such as the target sequence MAPK10 and other sequences, but not limited to the target gene.

The DNA molecule capable of encoding the oligonucleotide also belongs to the protection scope of the present invention. Furthermore, expression cassettes, recombinant vectors or recombinant bacteria containing said DNA molecules also belong to the protection scope of the present invention.

A second object of the invention is to provide an application.

The application provided by the present invention is specifically: the application of the oligonucleotide or the DNA molecule or the expression cassette, recombinant vector or recombinant bacteria in any of the following:

(a1) preparing a drug for inhibiting tumor growth, and/or inhibiting tumor growth;

(a2) preparing drugs for inhibiting tumor cell proliferation, and/or inhibiting tumor cell proliferation;

(a3) preparing a drug for inhibiting tumor cell invasion, and/or inhibiting tumor cell invasion;

(a4) preparing a drug for inhibiting tumor cell migration, and/or inhibiting tumor cell migration;

(a5) preparing a drug for promoting tumor cell apoptosis, and/or promoting tumor cell apoptosis;

(a6) Prepare a drug for preventing tumor recurrence, and/or prevent tumor recurrence.

Wherein, the prevention of tumor recurrence specifically refers to the prevention of tumor recurrence caused by residual tumor cells after tumor excision or radiotherapy and chemotherapy.

The third object of the present invention is to provide a medicine.

The medicine provided by the present invention has at least one of the following functions (a1)-(a6), and its active ingredient is the oligonucleotide or the DNA molecule or the expression cassette, recombinant vector or recombinant bacteria;

(a1) inhibiting tumor growth;

(a2) inhibit tumor cell proliferation;

(a3) inhibit tumor cell invasion;

(a4) inhibit tumor cell migration;

(a5) promote tumor cell apoptosis;

(a6) Prevention of tumor recurrence.

The medicament may also include a pharmaceutically acceptable carrier as needed. The pharmaceutically acceptable carrier should be compatible with the double-stranded RNA molecule in the medicament of the present invention.

Further, the pharmaceutically acceptable carrier can be an in vivo transfection reagent, such as liposome, polyethyleneimine (PEI), cationic nanopolymer and the like. Or the traditional drug carrier, sugar, starch, cellulose and its derivatives, vegetable oil isotonic buffer, etc. Or new drug carrier preparations: macromolecular substances, such as immunoglobulin, albumin, fibrin, glucose, etc.; cells that have been modified or not, such as white blood cells, red blood cells, etc.; synthetic biodegradable macromolecular lipids Body, such as intravenous milk, magnetic spheres, jade lipoglycan spheres, etc.; synthetic non-biodegradable macromolecules, such as semi-permeable microcapsules, polypropylene gel, etc. The medicament of the present invention can be made into various medically acceptable dosage forms, and can be given by doctors according to factors such as patient types, disease conditions, and administration methods and taking into account beneficial dosages for patients. Specific dosage forms may include various liquid dosage forms such as oral liquids, injections, and sublingual buccal agents, or various other dosage forms such as powders, tablets, and capsules by adding appropriate excipients. The preferred preparation maintains the performance of the oligonucleotide, has low toxicity and side effects, and is suitable for administration.

In an embodiment of the present invention, the pharmaceutically acceptable carrier is specifically liposome Lipofectamine2000 (Invitrogen). Correspondingly, in the drug, the ratio of the oligonucleotide to the Lipofectamine2000 is specifically (5-50) pmol: 1 μL, such as (20-32) pmol: 1 μL, specifically such as 20 pmol: 1 μL, or 30 pmol: 1 μL, or 50 pmol: 1 μL. More specifically, in the drug, the concentration of the oligonucleotide solution is 20 μmol/L, and the volume ratio between the oligonucleotide solution and the Lipofectamine2000 is specifically (1-1.6):1, such as 1:1 , or 1.5:1, or 1.6:1.

Of course, the drug may also include glucose, physiological saline or buffer solution and the like.

In the present invention, the tumor is specifically prostate cancer; the tumor cells are specifically prostate cancer cells. More specifically, the prostate cancer cells are DU-145 cells or PC-3 cells.

Experiments have proved that the oligonucleotide provided by the present invention inhibits the expression of MAPK10 in prostate cancer tumor cells, thereby achieving the effect of inhibiting tumor growth. At the animal level, experiments in which tumor cells were inoculated into nude mice and then treated with the oligonucleotide proved that the oligonucleotide can effectively inhibit tumor growth in vivo. In addition, at the cell level, it has also been confirmed that the oligonucleotide provided by the present invention has the functions of inhibiting tumor cell proliferation, inhibiting tumor cell invasion, inhibiting tumor cell migration and promoting tumor cell apoptosis. Therefore, the oligonucleotide provided by the present invention, as a novel small nucleic acid antitumor drug, can not only inhibit the growth of tumors in the body, reduce the risk of tumor spread, but also is expected to be used to prevent residual tumors after tumor resection or radiotherapy and chemotherapy. cells that cause tumor recurrence.

Description of drawings

Fig. 1 is a bar graph of inhibition of tumor cell proliferation detected by the 2'-OH oligonucleotide prepared in Example 1 by the CCK-8 method.

Fig. 2 is a bar graph showing that the 2'-OH oligonucleotide prepared in Example 1 inhibits tumor cell invasion.

FIG. 3 is a bar graph of inhibition of tumor cell migration by 2′-OH oligonucleotides prepared in Example 1. FIG.

FIG. 4 is a bar graph showing that 2′-OH oligonucleotides prepared in Example 1 promote tumor cell apoptosis.

Fig. 5 is the animal experiment results of treating prostate cancer with 2'-OH oligonucleotide prepared in Example 1. Among them, A is the 16th day after the injection of tumor cells (that is, the beginning of inoculation of tumor cells) is recorded as the 0th day, and the change of tumor volume of each group of mice over time; The 16th day after the tumor cell start) was recorded as the 0th day, and the tumor weight of each group of mice on the 14th day; C was recorded as the 0th day on the 16th day after the injection of the tumor cells (that is, the beginning of inoculation of the tumor cells) , Tumor photos of mice in each group on day 14.

detailed description

The experimental methods used in the following examples are conventional methods unless otherwise specified.

The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

For the quantitative experiments involved in the following examples, at least 3 repetitions were set, and the results were averaged.

5-week-old male nude mice: Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. (BALB/cNude401).

DU-145 cells: Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences (TCHu222).

PC-3 cells: Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences (TCHu158).

Embodiment 1, the preparation of oligomeric nucleic acid

The oligonucleotide described in the present invention is a double-stranded RNA; the double-stranded RNA is composed of a sense strand and an antisense strand, and the sequence of the sense strand is the sequence shown in sequence 1 in the sequence listing (5'-UUAUUGCUUAAGAAUACGCGUAG-3' ), the sequence of the antisense strand is the sequence shown in sequence 2 in the sequence listing (5'-CUACGCGUAUUCUUAAGCAAUAA-3'). A random sequence was selected as a negative control (NC). The sequence of the sense strand of NC is: 5'-UUCUCCGAACGUGUCACGUTT-3' (SEQUE 3), and the sequence of the antisense strand is: 5'-ACGUGACACGUUCGGAGAATT-3' (SEQUE 4). A, G, C, and U in the oligonucleotide and the negative control sequence represent adenine ribonucleotides, guanine ribonucleotides, cytosine ribonucleotides and uracil ribonucleotides, T Indicates thymidine nucleotide.

The oligonucleotide and the negative control were synthesized by Shanghai GenePharma Pharmaceutical Technology Co., Ltd. The oligonucleotide composed of sequence 1 and sequence 2 synthesized in the present invention is named 2'-OH-oligonucleotide.

Of course, the double-stranded RNA can also be replaced by a chimeric double strand formed by complementary pairing of a single-stranded RNA and a single-stranded DNA; the sequence of the single-stranded RNA in the chimeric double-strand is The sequence shown in sequence 1 (5'-UUAUUGCUUAAGAAUACGCGUAG-3'), the sequence of the single-stranded DNA is the sequence shown after replacing U in sequence 2 in the sequence listing with T (5'-CTACGCGTATTCTTAAGCAATAA-3');

Effect of embodiment 2, 2'-OH-oligonucleotide on tumor cell proliferation

The 2′-OH oligonucleotide prepared in Example 1 and the negative control powder were respectively dissolved in DEPC water to prepare a solution with a final concentration of 20 μmol/L. Take logarithmic phase prostate cancer cells (DU-145 cells and PC-3 cells), adjust the concentration of the cell suspension to 5×10 ⁵ cells/ml, inoculate 100 μl per well in a 96-well plate; place at 37°C, The cells were allowed to adhere to the wall in an incubator containing 5% CO ₂ and cultured overnight. For transfection, dilute 0.3 μl of 2′-OH oligonucleotide solution (concentration: 20 μmol/L) and 0.3 μl of lipofectamine2000 (Invitrogen) in 25 μl of serum-free medium (Opti-MEM), mix well, and incubate at room temperature 5 minutes, then the above-mentioned oligonucleotide solution was mixed with lipofectamine2000 solution, left standing at room temperature for 20 minutes, and about 50 μl of oligonucleotide-liposome complex was added to the cell culture plate (the cell growth fusion rate was 65% ), respectively cultured for 0, 24, 48, 72 and 96 hours; carefully sucked off the supernatant, added 90 μl of fresh medium, and then added 10 μl of CCK-8, continued to cultivate for 2.5 hours, and measured the concentration of each well at 490 nm in an enzyme-linked immunosorbent assay Absorbance value; at the same time, set zero-adjustment wells (medium, CCK-8), and set 5 duplicate wells for each group. The larger the value of OD490, the more the number of living cells, that is, the stronger the cell proliferation ability; on the contrary, the smaller the value of OD490, the less the number of living cells, that is, the weaker the cell proliferation ability.

The results are shown in Figure 1, which shows that the proliferation of tumor cells transfected with the 2'-OH oligonucleotide prepared in Example 1 was significantly inhibited, and compared with the negative control group (NC), the proliferation rate was significantly reduced.

Example 3, 2'-OH-oligonucleotide inhibits tumor cell invasion ability

The 2′-OH oligonucleotides prepared in Example 1 were subjected to transfection experiments according to the grouping and experimental system of Example 2. After the transfection was completed, they were cultured overnight at 37° C. in an incubator containing 5% CO ₂ , and replaced with Culture medium containing 5% (volume fraction) FBS was starved for 24 hours. Before the experiment, dilute 50 mg/L Matrigel (Matrigel ^TM product number 354234 of BD Company) 1:6 with DMEM culture solution containing 0.5% (volume fraction) FBS, take 60 μl of the upper chamber surface coated with the bottom membrane of the transwell chamber, and place it at 37°C for 5 Incubate for 4 hours in a % CO ₂ incubator and discard the supernatant. The transfected prostate cancer cells PC-3 and DU145 were digested with trypsin, counted on a hemocytometer, inoculated in the transwell chamber at a concentration of 5×10 ³ cells/well, and 700 μl of medium with a serum concentration of 20% was added to the lower chamber. Incubate at 37°C with 5% CO ₂ for 48h. Take out the chamber and wash once with PBS. Add pre-cooled methanol -20 ℃ fixed 10min. Wash with PBS three times after discarding the methanol. Gently scrape off the Matrigel glue on the upper surface of the chamber with a cotton swab, and wash the upper surface 3 times with PBS. Take the chamber upside down and air dry. Put the chamber into a new 24-well, add 200 μl of 0.1% crystal violet (preparation method: weigh 0.1 g of crystal violet and add 100 ml of glacial acetic acid to dissolve completely, shake well), submerge the membrane, and stain at 37°C for 30 minutes. Wash ₃ times with ddH2O. After the chamber is air-dried, put it into the well, take several fields of view under an inverted microscope, take pictures and count, and count the number of cells passing through the membrane. The more tumor cells that pass through the membrane, the stronger the invasive ability of the tumor cells; on the contrary, the fewer the number of tumor cells that pass through the membrane, the weaker the invasive ability of the tumor cells.

The results are shown in Figure 2. Figure 2 shows that compared with the negative control group (NC), the invasion ability of the tumor cells transfected with the 2'-OH oligonucleotide prepared in Example 1 was significantly inhibited.

Example 4, 2'-OH-oligonucleotide inhibits tumor cell migration ability.

The 2′-OH oligonucleotide prepared in Example 1 and the negative control powder were respectively dissolved in DEPC water to prepare a solution with a final concentration of 20 μmol/L. Prostate cancer cells in logarithmic phase (DU-145 cells and PC-3 cells) were taken, and the concentration of the cell suspension was adjusted to 1.5×10 ⁶ cells/ml. Inoculate a 6-well plate at a concentration of 1.5×10 ⁶ cells/well, mix well and incubate overnight at 37°C in 5% CO ₂ . For transfection, dilute 8 μl of 2′-OH oligonucleotide solution (concentration: 20 μmol/L) and 5 μl of lipofectamine2000 (Invitrogen) in 250 μl of serum-free medium (Opti-MEM), mix well, and incubate at room temperature for 5 minutes , then the above-mentioned oligonucleotide solution was mixed with lipofectamine2000 solution, left at room temperature for 20 minutes, and about 500 μl of oligonucleotide-liposome complex was added to the cell culture plate (cell growth fusion rate was 85%). After transfection, the 6-well plate was incubated overnight at 37°C in 5% CO ₂ , and a 20 μl pipette tip was compared with a ruler, and a straight line was evenly drawn across the wells in the 6-well plate, with three stripes per well. The cells were washed 3 times with PBS, the scratched cells were removed, and 3% (volume fraction) FBS medium was added. Place them in a 5% CO ₂ incubator at 37°C to continue culturing, and take pictures at 0h, 8h, 24h, and 48h respectively. Use Image-ProPlus to measure the area and length of the scratches in each photo, and calculate the width of each scratch according to the scratch width value = scratch area/scratch length value; then calculate the scratch healing rate at the 48th hour, and at the 48th hour Scratch healing rate=(0h scratch width-48h scratch width)/0h scratch width. The greater the scratch healing rate, the stronger the migration ability of tumor cells; on the contrary, the smaller the scratch healing rate, the weaker the migration ability of tumor cells.

The results are shown in Figure 3, which shows that, compared with the negative control group (NC), the migration ability of the tumor cells transfected with the 2'-OH oligonucleotide prepared in Example 1 was significantly inhibited.

Example 5, 2'-OH-oligonucleotide promotes tumor cell apoptosis.

The 2'-OH oligonucleotides prepared in Example 1 were subjected to transfection experiments according to the grouping and experimental system of Example 4. After transfection, culture at 37°C in an incubator containing 5% CO ₂ for 48 hours, digest the cells with EDTA-free trypsin, collect the cells by centrifugation, and centrifuge at 2000 rpm for 5 minutes, then discard the medium. Wash the cells twice with cold PBS (2000rpm, centrifuge for 5min to collect the cells).

The cells were resuspended with 400 μl 1×BindingBuffer (MbchemM3036), and the cell concentration was adjusted to 1×10 ⁶ cells/ml. Add 5 μl AnnexinV-FITC to the cell suspension, mix gently and incubate at 2-8°C for 15 minutes in the dark. After adding 10 μl PI, mix gently and incubate at 2-8°C in the dark for 5 minutes. within 1 hour by flow cytometry. Normal cells with no fluorescence or very low background fluorescence; early apoptotic cells with only strong green fluorescence; late apoptotic cells with red and green double fluorescence; necrotic cells with only strong red fluorescence .

The results are shown in Figure 4, and Figure 4 shows that, compared with the negative control group (NC), the apoptotic ratio of the tumor cells transfected with the 2'-OH oligonucleotide prepared in Example 1 was significantly higher, i.e. 1 The prepared 2'-OH oligonucleotide can promote the apoptosis of tumor cells.

Inhibitory effect of embodiment 6, 2'-OH oligonucleotide on tumor growth

Prostate cancer cells (DU-145 cells and PC-3 cells) in the logarithmic growth phase were taken, and single-cell suspensions were prepared respectively, and the cell concentration was adjusted to 7×10 ⁷ cells/ml. Inject 0.2ml of cell suspension in the upper part of the groin of male nude mice at the age of 5 weeks, and inject 10 of them altogether. After 16 days of injection, subcutaneous solid tumors of ⁵ mm can be seen. The 2'-OH oligonucleotide prepared in Example 1 and Negative control treatment. The nude mice were divided into two groups, one group was treated with 2′-OH oligonucleotide, and the other group was negative control group. The method is as follows: Dissolve the 2′-OH oligonucleotide and the negative control in RNase-free sterile water to prepare a solution with a final concentration of 20 μmol/L. Using lipofectamine2000 (Invitrogen) as the transfection reagent, 150 μl of the oligonucleotide Polynucleic acid solution (concentration: 20 μmol/L) and 100 μl lipofectamine2000 (Invitrogen) were diluted in 150 μl serum-free medium (Opti-MEM) and mixed, incubated at room temperature for 5 minutes, and then the above oligonucleotide solution was mixed with lipofectamine2000 solution , after standing at room temperature for 20 minutes, insert the needle about 5mm away from the tumor, subcutaneously sneak to the tumor, inject slowly at multiple points, 100 μl per nude mouse, inject once every two days, and inject 5 times in total. After three days of feeding, all nude mice were sacrificed, and the tumors were taken out and weighed. Sixteen days after the injection of tumor cells (that is, the start of tumor cell inoculation), the length and short axis of the tumor were measured every two days, and the volume of the tumor was calculated. The calculation formula is: V(mm ³ )=0.5×long diameter×short diameter ² .

The results are shown in Figure 5, showing that on the tumor model of prostate cancer, the volume and weight of nude mouse tumors treated with 2'-OH oligonucleotides were significantly smaller than the negative control treatment group (NC), indicating that the 2'-OH Oligonucleotides can significantly inhibit the growth of tumors.

Claims (10)

1. a nucleic acid oligomer, for as follows (A) or (B):

(A) double-stranded RNA: described double-stranded RNA is made up of positive-sense strand and antisense strand, described positive-sense strand has sequence shown in sequence 1 in sequence table, and described antisense strand has sequence shown in sequence 2 in sequence table；

(B) the chimeric double-strand formed by a single stranded RNA and a single stranded DNA complementary pairing: the described single stranded RNA in described chimeric double-strand has sequence shown in sequence 1 in sequence table, described single stranded DNA have the U in sequence in sequence table 2 is replaced with after T shown in sequence.

2. nucleic acid oligomer according to claim 1, it is characterised in that: in described (A), the sequence of described positive-sense strand is sequence 1 in sequence table, and described antisense strand is sequence 2 in sequence table；

In described (B), the sequence of described single stranded RNA is sequence 1 in sequence table, and the sequence of described single stranded DNA is the sequence formed after the U in sequence in sequence table 2 is replaced with T.

3. nucleic acid oligomer according to claim 1 and 2, it is characterised in that: described nucleic acid oligomer at least one in following modification:

(1) modification to the phosphodiester bond connecting nucleotide in the nucleotide sequence of described nucleic acid oligomer；

(2) modification to 2 '-OH of the ribose in the nucleotide sequence of described nucleic acid oligomer；

(3) modification to the base in the nucleotide sequence of described nucleic acid oligomer.

4. the DNA molecular of nucleic acid oligomer described in coding claim 1 or 2.

5. contain the expression cassette of DNA molecular described in claim 4, recombinant vector or recombinant bacterium.

6. the DNA molecular described in arbitrary described nucleic acid oligomer or claim 4 or the expression cassette described in claim 5, recombinant vector or the recombinant bacterium application in arbitrary as follows in claim 1-3:

(a1) preparation is for suppressing the medicine of tumor growth, and/or suppresses tumor growth；

(a2) preparation is for suppressing the medicine of tumor cell proliferation, and/or suppresses tumor cell proliferation；

(a3) preparation is for suppressing the medicine of tumor cell invasion, and/or suppresses tumor cell invasion；

(a4) preparation is for suppressing the medicine of tumor cell migration, and/or suppresses tumor cell migration；

(a5) preparation is for promoting the medicine of apoptosis of tumor cells, and/or promotes apoptosis of tumor cells；

(a6) preparation is for the medicine of prophylaxis of tumours recurrence, and/or prophylaxis of tumours recurrence.

7. having medicine at least one in following (a1)-(a6) function, its active component is the DNA molecular described in arbitrary described nucleic acid oligomer or claim 4 or the expression cassette described in claim 5, recombinant vector or recombinant bacterium in claim 1-3；

(a1) tumor growth is suppressed；

(a2) tumor cell proliferation is suppressed；

(a3) tumor cell invasion is suppressed；

(a4) tumor cell migration is suppressed；

(a5) apoptosis of tumor cells is promoted；

(a6) prophylaxis of tumours recurrence.

8. medicine according to claim 7, it is characterised in that: described medicine also includes pharmaceutically acceptable carrier.

9. medicine according to claim 8, it is characterised in that: described pharmaceutically acceptable carrier is Lipofectamine2000；In described medicine, the proportioning of described nucleic acid oligomer and described Lipofectamine2000 is (5-50) pmol:1 μ L.

10. arbitrary described medicine in application according to claim 6 or claim 7-9, it is characterised in that: described tumor is carcinoma of prostate；Described tumor cell is prostate gland cancer cell.