The present invention relates to a pharmaceutical composition for cancer therapy comprising a transcription or processing product of a gene encoding a particular miRNA.
MicroRNA (hereinafter miRNA) is a functional nucleic acid that is encoded on the genome and ultimately becomes a minuscule RNA of about 20-25 bases long through a multistep production process. miRNA is classified as a functional ncRNA (non-coding RNA), and it is being elucidated that it plays an important role in various biological phenomena (such as regulation of gene expression etc.). Various miRNAs that has come to be well-known thus far including human miRNA are registered in the miRBase (see http://www.mirbase.org/).
miRNA is indicated to be associated with the onset and progression of e.g. cancer, cardiovascular disease, neurodegenerative disease, psychiatric disease, chronic inflammatory disease, and the like. Particularly in recent years, it has been indicated that miRNA is deeply involved in canceration or aging of cells.
For example, Patent Literature 1 describes that miR-22 promotes the aging of cells and suppresses invasion and metastasis of cancer. Moreover, Patent Literature 2 describes that a composition comprising miR-34 may be employed for cancer therapy.
The present inventors diligently searched for miRNAs having therapeutic effect against cancer from among a great number of miRNAs.
As a result, the present inventors found particular miRNAs that have significantly stronger cancer therapeutic effect than the miRNAs described in prior art, thus arriving at the completion of the present invention.
In other words, the present invention relates to a pharmaceutical composition for cancer therapy comprising a transcription or processing product of a gene encoding a miRNA, characterized in that said miRNA is one or more miRNAs selected from the group consisting of miR-3140, miR-137, miR-631, and miR-657.
One embodiment of the present invention is characterized in that said cancer is a solid cancer.
One embodiment of the present invention is characterized in that said solid cancer is colon cancer, pancreatic cancer, tongue cancer, mesothelioma, uterine sarcoma, osteosarcoma, breast cancer, lung cancer, or head and neck cancer.
One embodiment of the present invention is characterized in that said transcription or processing product of a gene encoding a miRNA is a pri-miRNA, a pre-miRNA, a double-stranded mature-miRNA, a single-strand mature-miRNA expressed from the 5′-end of a pre-miRNA, or a single-strand mature-miRNA expressed from the 3′-end of a pre-miRNA.
One embodiment of the present invention is characterized in that said miRNA is:
One embodiment of the present invention is characterized in that said miRNA is chemically modified.
One embodiment of the present invention is characterized in that said chemical modification is one or more chemical modifications selected from the group consisting of LNA-tion, BNA-tion, ENA-ation, 2′-OMe modification, phosphorothioation, S-TuD-ation, morpholino modification, peptide addition, glycosylation, aptamer addition, hydrophobic molecule addition, polymer addition, and addition of unmodified DNA.
One embodiment of the present invention is characterized in that said pharmaceutical composition further comprises a nucleic acid transfection agent.
One embodiment of the present invention is characterized in that said transfection agent is a lipid-based transfection agent, a polymer-based transfection agent, a magnetic particle-based transfection agent, an exosome for nucleic acid delivery, or a viral protein for nucleic acid delivery.
One embodiment of the present invention is characterized in that said transfection agent is a transfection agent comprising a peptide represented by amino acid sequences GGGGDD (G4D2), GGGGGGDD (G6D2), GGGGGGGGDD (G8D2), GGGGGGGGGGDD (G10D2), AAAAAAD (A6D), AAAAAADD (A6D2), AAAAAAK (A6K), AAAAAAKK (A6K2), VVVVVVD (V6D), VVVVVVDD (V6D2), VVVVVVK (V6K), VVVVVVKK (V6K2), LLLLLLD (L6D), LLLLLLDD (L6D2), LLLLLLK (L6K), or LLLLLLKK (L6K2).
One embodiment of the present invention is characterized in that the pharmaceutical composition of the present invention is for topical administration.
One embodiment of the present invention is characterized in that the pharmaceutical composition of the present invention is used in combination with other anticancer agents.
One embodiment of the present invention is characterized in that said other anticancer agents are one or more anticancer agents selected from the group consisting of an alkylating agent, a platinum preparation, a metabolism antagonist, a topoisomerase inhibitor, a microtubular inhibitor, an anti-cancerous antibiotic, a molecular target drug, a hormone preparation, an immunomodulation drug, an interferon, an interleukin, a plant-derived anticancer agent, and a BRM preparation.
Another embodiment of the present invention relates to the use of a transcription or processing product of a gene encoding a miRNA for manufacturing a pharmaceutical composition for cancer therapy, characterized in that said miRNA is one or more miRNAs selected from the group consisting of miR-3140, miR-137, miR-631, and miR-657.
Another embodiment of the present invention relates to a cancer therapy method comprising a step of applying to a cancer patient a therapeutically effective amount of an anti-cancerous pharmaceutical composition comprising a transcription or processing product of a gene encoding a miRNA, characterized in that said miRNA is one or more miRNAs selected from the group consisting of miR-3140, miR-137, miR-631, and miR-657.
An invention of any combination of one or more characteristics listed above is encompassed by the scope of the present invention.
The present invention relates to a pharmaceutical composition for cancer therapy comprising a transcription or processing product of a gene encoding a miRNA (micro-RNA). A general miRNA is biosynthesized via a continuous process. The primary transcription product of a gene encoding a miRNA is called a Primary miRNA transcript (pri-miRNA), and generally has a stem-loop hairpin structure. Pri-miRNA is cleaved by a microprocessor complex, takes a hairpin form by Drosha which is a RNase III-series enzyme, and precursor miRNA (pre-miRNA) which is an intermediate precursor of about 70 bases is produced. The pre-miRNA is then transported from the nucleus to the cytoplasm. In the cytoplasm, it is further cleaved by Dicer which is another RNase III enzyme, and a double-stranded mature miRNA is produced. In general, among the two strands, “-5p” is added to that expressed from the 5′-end of the precursor and “-3p” is added to that expressed from the 3′-end, and are represented as “hsa-miR-21-5p” and “hsa-miR-21-3p”. Note that in principle, well-known miRNAs are registered in the miRBase (http://www.mirbase.org/).
Note that only one of the strands in the mature miRNA may exert the desired effect, or each of the stands may exert the desired effect, or the desired effect may be exerted in the double-stranded state. Moreover, the desired effect may also be exerted in pri-miRNA state or pre-miRNA state.
The nucleic acid comprised in the composition of the present invention may be a transcription or processing product of a gene encoding one or more miRNAs selected from the group consisting of miR-3140, miR-137, miR-631, and miR-657, and may also be a variant or a modification that retains the function of the aforementioned nucleic acid.
The sequence of the transcription or processing product of the gene encoding miR-3140 used in one embodiment of the present invention is as follows.
Note that the sequence of the intron region encoding the pri-miRNA of miR-3140 is shown in SEQ ID NO. 4.
The sequence of the transcription or processing product of the gene encoding miR-137 used in one embodiment of the present invention is as follows.
Note that the sequence of the intron region encoding the pri-miRNA of miR-137 is shown in SEQ ID NO. 7.
The sequence of the transcription or processing product of the gene encoding miR-631 used in one embodiment of the present invention is as follows.
Note that the sequence of the intron region encoding the pri-miRNA of miR-631 is shown in SEQ ID NO. 10.
The sequence of the transcription or processing product of the gene encoding miR-657 used in one embodiment of the present invention is as follows.
Note that the sequence of the intron region encoding the pri-miRNA of miR-657 is shown in SEQ ID NO. 13.
The nucleic acid comprised in the composition of the present invention may be e.g. a nucleic acid having substitution, addition, and/or deletion of 1, 2, 3, 4, or 5 bases to a mature-miRNA consisting of a sequence represented by SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 5, SEQ ID NO. 8, or SEQ ID NO. 11, as well as having the effect of inhibiting cancer cell growth. The substitution of a base to the mature-miRNA may be e.g. a conservative substitution of RNA known in the art.
Moreover, the nucleic acid comprised in the composition of the present invention may be e.g. a nucleic acid having 80% or more (preferably, 85% or more, 90% or more, 95% or more) sequence homology (or sequence identity) to a mature-miRNA consisting of a sequence represented by SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 5, SEQ ID NO. 8, or SEQ ID NO. 11, as well as having the effect of inhibiting cancer cell growth.
A mature-miRNA consisting of a sequence represented by SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 5, SEQ ID NO. 8, or SEQ ID NO. 11 can be easily manufactured by a RNA synthesis equipment commonly used in the art, and a nucleic acid having a particular base substituted, added, and/or deleted can similarly be easily manufactured. Moreover, numerous companies accept commissioned synthesis of nucleic acids, and it is also easy to obtain RNA of the desired sequence from such companies. Accordingly, those skilled in the art will be able to investigate the nature and function of a variant or modification of a mature-miRNA consisting of a sequence represented by SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 5, SEQ ID NO. 8, or SEQ ID NO. 11 by conventional means without excessive burden.
Moreover, the nucleic acid comprised in the composition of the present invention may have received a chemical modification well-known in the art with the purpose of improving the stability or specificity etc. of RNA. Chemical modification that may be employed in the present invention may be e.g. LNA (Locked Nucleic Acid)-tion, BNA (Bridged Nucleic Acid)-tion, ENA (2′-O,4′-C-Ethylene-bridged Nucleic Acids)-tion, 2′-OMe modification, phosphorothioation (S-ation), S-TuD (Synthetic Tough Decoy)-ation, morpholino modification, peptide addition, glycosylation, aptamer addition, hydrophobic molecule addition such as cholesterol, polymer addition such as PEG, or addition of unmodified DNA. These chemical modifications can be performed by a well-known means known in the art.
The present invention can be employed for various cancer therapies, e.g. it can be favorably employed for solid cancer. More preferably, the subject of the present invention may be colon cancer, pancreatic cancer, tongue cancer, mesothelioma, uterine sarcoma, osteosarcoma, breast cancer, lung cancer, or head and neck cancer.
In order to appropriately introduce the present invention to cancer cells, the composition of the present invention may further comprise a nucleic acid transfection agent. Examples of the nucleic acid transfection agent that may be employed for the present invention include a lipid-based transfection agent, a polymer-based transfection agent, a magnetic particle-based transfection agent, an exosome for nucleic acid delivery, or a viral protein for nucleic acid delivery.
An example of the lipid-based transfection agent includes a cationic lipid. With a cationic lipid, nucleic acid-cationic lipid complexes are incorporated into cells via endocytosis and released into the cytoplasm, thus introducing the nucleic acid into cells (lipofection). Specifically, e.g. various commercially available reagents for lipofection may be employed.
An example of the polymer-based transfection agent includes e.g. a cationic polymer. When a cationic polymer comes in contact with a nucleic acid, a nucleic acid-polymer complex is formed, and the complex attaches to the cell membrane via electrostatic interaction and is incorporated into the cell via endocytosis. Specifically, a cationic peptide and a derivative thereof (such as polylysine and polyornithine), a linear or branched-chain synthetic polymer (such as polybrene and polyethyleneimine), a polysaccharide-based introduction molecule (such as cyclodextrin and chitosan), a natural polymer (such as histone and collagen), as well as active and inactive dendrimers, and the like may be employed. Moreover, transfection agents employed in so-called nanoDDS, such as a transfection agent that employs a block copolymer that forms micellar nanoparticles and a transfection agent that employs carbon nanohorns can also be used for the present invention.
An example of the magnetic particle-based transfection agent includes a transfection agent that employs magnetic particles coated with cation molecules. A magnetic particle-based transfection agent carries out transfection by adhering the nucleic acid on the surface of magnetic particles, and then magnetically introducing the aforementioned magnetic particles into cells. Specifically, for example various commercially available magnetic particles for transfection may be employed.
Moreover, nucleic acid transfection agent that employs a generally available exosome or a transfection agent that utilizes viral proteins such as adenovirus can also be used for the present invention.
Further, in the present invention, a transfection agent comprising a peptide represented by amino acid sequences GGGGDD (G4D2) (SEQ ID NO. 14), GGGGGGDD (G6D2) (SEQ ID NO. 15), GGGGGGGGDD (G8D2) (SEQ ID NO. 16), GGGGGGGGGGDD (G10D2) (SEQ ID NO. 17), AAAAAAD (A6D) (SEQ ID NO. 18), AAAAAADD (A6D2) (SEQ ID NO. 19), AAAAAAK (A6K) (SEQ ID NO. 20), AAAAAAKK (A6K2) (SEQ ID NO. 21), VVVVVVD (V6D) (SEQ ID NO. 22), VVVVVVDD (V6D2) (SEQ ID NO. 23), VVVVVVK (V6K) (SEQ ID NO. 24), VVVVVVKK (V6K2) (SEQ ID NO. 25), LLLLLLD (L6D) (SEQ ID NO. 26), LLLLLLDD (L6D2) (SEQ ID NO. 27), LLLLLLK (L6K) (SEQ ID NO. 28), or LLLLLLKK (L6K2) (SEQ ID NO. 29) may be employed, and in particular AAAAAAD (A6D) or AAAAAAK (A6K) can be favorably employed. The effects of these peptides as transfection agents are disclosed in e.g. WO2010/024262.
The pharmaceutical composition for cancer therapy of the present invention can be used in combination with other anticancer agents well-known in the art. Other anticancer agents used in combination are not limited, and e.g. one or more anticancer agents selected from the group consisting of an alkylating agent, a platinum preparation, a metabolism antagonist, a topoisomerase inhibitor, a microtubular inhibitor, an anti-cancerous antibiotic, a molecular target drug, a hormone preparation, an immunomodulation drug, an interferon, an interleukin, a plant-derived anticancer agent, and a BRM preparation can be employed.
The aspects of combination use of the pharmaceutical composition for cancer therapy of the present invention and the other anticancer agents well-known in the art are not limited, and can be carried out by those skilled in the art (such as a physician) in various aspects according to the type of cancer to be the subject or therapeutic stage and the like. The pharmaceutical composition for cancer therapy of the present invention and the other anticancer agents may be administered to the subject at the same or different times. The pharmaceutical composition for cancer therapy of the present invention and the other anticancer agents may also be prepared as formulations comprising each and administered to a subject. The pharmaceutical composition for cancer therapy of the present invention and the other anticancer agents may also be prepared as a kit that separately comprises each.
An aspect of administering the pharmaceutical composition for cancer therapy of the present invention and the other anticancer agents at the same time may be e.g. an aspect of administering to a subject a formulation comprising the pharmaceutical composition for cancer therapy of the present invention and the other anticancer agents.
In the present invention, an aspect of administering the pharmaceutical composition for cancer therapy of the present invention and the other anticancer agents at different times may be e.g. an aspect of administering each of the pharmaceutical composition for cancer therapy of the present invention and the other anticancer agents with staggered time, and e.g. an aspect of administering the pharmaceutical composition for cancer therapy of the present invention and the other anticancer agents from different administration routes.
The administration route of the pharmaceutical composition for cancer therapy of the present invention is not limited, and may be systemic administration or topical administration. Administration routes can include e.g. oral administration including sublingual administration, parenteral administration such as inhalation administration, direct administration to target tissue by catheter or injection, intravenous administration including infusion, transdermal administration by patches and the like, suppositories, or administration by forced enteral nutrition employing a nasogastric tube, a nasointestinal tube, a gastrostomy tube, or enterostomy tube, and the like.
The dosage form of the pharmaceutical composition for cancer therapy of the present invention may be appropriately determined according to said administration route, and can include, but is not limited to, injections, infusions, tablets, capsules, fine granules, powders, liquids, solutions dissolved in syrups etc., patches, suppositories, and the like.
The subject for administering the pharmaceutical composition of the present invention is not limited, and e.g. the present invention can be employed for mammals (humans, pigs, cows, monkeys, baboons, dogs, cats, rats, mice, and the like). However, when it is unfavorable, humans can be removed from subjects.
The administration method of the pharmaceutical composition of the present invention to a subject (administration route, dosage, administration frequency per day, administration timing, and the like) is not limited, and can be appropriately determined by those skilled in the art (such as a physician) according to the health state of the subject, the extent of disease, the type of agent used in combination, and the like.
The terms used herein, except for those that are particularly defined, are employed for describing particular embodiments, and do not intend to limit the invention.
Moreover, the term “comprising” as used herein, unless the content clearly indicates to be understood otherwise, intends the presence of the described items (such as components, steps, elements, and numbers), and does not exclude the presence of other items (such as components, steps, elements, and numbers).
Unless otherwise defined, all terms used herein (including technical and scientific terms) have the same meanings as those broadly recognized by those skilled in the art of the technology to which the present invention belongs. The terms used herein, unless explicitly defined otherwise, are to be construed as having meanings consistent with the meanings herein and in related technical fields, and shall not be construed as having idealized or excessively formal meanings.
The present invention will now be described in further detail with reference to Examples. However, the present invention can be embodied by various aspects, shall not be construed as being limited to the Examples described herein.
(1) Transfection of all miRNAs into TIG-3 Cells (
A total of 2028 types of miRNAs (mirVana; Ambion) were transfected into TIG-3 cells (human fibroblasts) with an auto dispenser Bravo (Agilent). Transfection was carried out by the following protocol.
Setting the day of transfection as Day 0, staining was performed five days later in order to evaluate the number of cells and cell size. The protocol therefor is shown below.
(2-2) SA-β-Galactosidase Assay of Transfected Cells
Setting the day of transfection as Day 0, SA-β-galactosidase assay was performed on Day 7. The operating protocol is shown below.
(3) Analysis of the Screening Result (
Each of the values of the number of cells and cell size obtained were scored with the numeric values of random sequence miRNA which is the negative control and the numeric values of miR-34a-Sp which is the positive control. The scoring method is shown below.
Within Each Plate
The 349 types of aging induction miRNA (mirVana; Ambion) obtained from screening were transfected into various cancer cell strains (large intestine cancer cell strain HCT116 (p53 wildtype and p53 deletion), pancreatic cancer cell strains BxPC-3 and CFPAC-1, and tongue cancer cell strain HSC-4) with an auto dispenser Bravo (Agilent).
Transfection was carried out with the following protocol.
Setting the day of transfection as Day 0, on Day 5 the cell survival rate was evaluated with PrestoBlue (Invitrogen). The operating protocol is shown below.
The above protocol was carried out for each cancer cell strain (large intestine cancer cell strain HCT116 (p53 wildtype and p53 deletion), pancreatic cancer cell strains BxPC-3 and CFPAC-1, and tongue cancer cell strain HSC-4), and the miRNAs that suppress cell proliferation more significantly than miR-34a-5p were sorted.
As a result of sorting the miRNAs that suppress cell proliferation more significantly than miR-34-5p commonly in all cell strains, four miRNAs (miR-137-3p, miR-631-5p, miR-657-3p, and miR-3140-3p) were found (
(1) Tongue Cancer Cell Strain (HSC-4 and OSC-19) (
Four miRNAs (miR-137-3p, miR-631-5p, miR-657-3p, and miR-3140-3p) were transfected into tongue cancer cell strains (HSC-4 and OSC-19), and cell proliferation was observed. Transfection was carried out with the following protocol.
As shown in
(2) Tongue Cancer Cell Strain (SAS) (
Four miRNAs (miR-137-3p, miR-631-5p, miR-657-3p, and miR-3140-3p) were transfected into a tongue cancer cell strain (SAS), and cell proliferation was observed. Transfection was carried out with the following protocol.
As shown in
[Table 7]
(3) Malignant Pleural Mesothelioma Cell Strains, Uterine Sarcoma Cell Strain, and Osteosarcoma Cell Strain (
Among the above four miRNAs, miR-3140-3p which had particularly high cell growth inhibition effect was transfected in each of malignant pleural mesothelioma cell strains (MSTO-211H and EHMES-10), uterine sarcoma cell strain (MES-SA), and osteosarcoma cell strain (U2-OS), and cell proliferation was observed. As comparison subjects, miR-22-3p and miR-34a-5p which showed cell growth inhibition effect in prior research were employed. Transfection was carried out with the following protocol.
As shown in
(4) Breast Cancer Stem Cell Strain (MDA-MB231-Luc-D3H2LN Cells) (
miR-3140-3p was transfected in a highly metastatic cancer cell strain of breast cancer (MDA-MB231-luc-D3H2LN cells), and cell proliferation was observed. As comparison subjects, miR-22-3p and miR-34a-5p which showed cell growth inhibition effect in prior research were employed. Transfection was carried out with the following protocol.
Moreover, by the protocol below, the expression of apoptosis marker Annexin V in breast cancer cells transfected with miR-3140-3p was analyzed.
As shown in
Moreover, as shown in
(5) Lung Cancer Cell Strain (A549) (
miR-3140-3p was transfected into lung cancer cell strain (A549), and cell proliferation was observed. As comparison subjects, miR-22-3p and miR-34a-5p which showed cell growth inhibition effect in prior research were employed. Transfection was carried out with the following protocol.
As shown in
(6) Large Intestine Cancer Cell Strains (SW620, SW480, and HT29) (
miR-3140-3p was transfected into large intestine cancer cell strains (SW620, SW480, and HT29), and cell proliferation was observed. Transfection was carried out with the following protocol.
Results are shown in
In order to verify the antitumor effect of the miRNA of the present invention in vivo, experiments was performed with laboratory animals transplanted with malignant pleural mesothelioma cells.
(1) Preparation of Cells
Malignant pleural mesothelioma cells MSTO-211H cells were used.
Six weeks-old C-B-17/Icr-scid/scid Jel (SCID mouse) were used as mice. SCID mice were subcutaneously administered 100 μL of the prepared cell suspension, and the cells were allowed to settle
(3) miRNA Administration
Negative control which is the control sequence and miR-3140-3p which is the miRNA of the present invention were employed. A6K (from 3D Matrix) was employed as the nucleic acid delivery reagent. The administration of miRNA to mice was carried out with the following protocol.
The experimental results were evaluated with the following protocol.
Experimental results are shown in
(1) Preparation of Cells
Malignant pleural mesothelioma cells EHMES-10 cells were used.
Six weeks-old C-B-17/Icr-scid/scid Jel (SCID mouse) were used as mice. SCID mice were subcutaneously administered 100 μL of the prepared cell suspension, and the cells were allowed to settle.
(3) miRNA Administration
Negative control which is the control sequence and miR-3140-3p which is the miRNA of the present invention were employed. A6K (from 3D Matrix) was employed as the nucleic acid delivery reagent. The administration of miRNA to mice was carried out with the following protocol.
The experimental results were evaluated with the following protocol.
Imaging analysis results of tumor tissues at 12, 19, 26, and 33 days after tumor cell transplantation are shown in
(1) Preparation of Cells
Tongue cancer cell strain HSC-4 cells were used.
Six weeks-old C-B-17/Icr-scid/scid Jcl (SCID mouse) were used as mice. SCID mice were subcutaneously administered 100 μL of the prepared cell suspension, and the cells were allowed to settle.
(3) miRNA Administration
Negative control which is the control sequence and miR-3140-3p, miR-137, miR-631 and miR-657 which are the miRNAs of the present invention were employed. A6K (from 3D Matrix) was employed as the nucleic acid delivery reagent. The administration of miRNA to mice was carried out with the following protocol.
The experimental results were evaluated with the following protocol.
Imaging analysis results of tumor tissues after tumor cell transplantation are shown in
From the above results, it was shown that the miRNA of the present invention also exerts extremely strong antitumor effect in vivo.
The tumor suppression effect of miR-3140-3p in vivo was investigated with intrathoracic orthotopic transplantation model mouse.
Six weeks-old male mice (C-B-17/Icr-scid/scid Jcl) were used as mice. Malignant pleural mesothelioma cell strain EHMES-10 which expresses the luciferase gene was used as the tumor cell.
The experiment protocol is shown below.
Administration and imaging schedule is shown in
Note that in order to improve expression efficiency, miRNA was administered as a double-strand in combination with a complementary strand comprising partial mismatch.
Experimental results are shown in
In the group with one administration of miR-3140-3p, tumor reduction was seen by the second imaging (Day 10 after transplantation), and the effect had persisted until the fourth imaging (Day 24 after transplantation) (
The imaging results were digitized and graphed, and it was shown that miR-3140-3p significantly suppressed tumor expansion (
By comparing the survival rates of mice, it was observed that the survival rates of mice had improved in the miR-3140-3p administration group (
It was shown that similarly to the group with one administration, miR-3140-3p also significantly suppressed malignant pleural mesothelioma in the groups with two and three administrations of miR-3140-3p (
The survival rates of mice were more improved in the group with three administrations of miR-3140-3p (
The IC50 value of miR-3140-3p was calculated with malignant pleural mesothelioma cell EHMES-10.
The protocol therefor is shown below.
Note that the same control RNA as that used in Experiment 7 was employed as the control.
Results are shown in
In order to investigate the relative superiority of antitumor effect of miR-3140-3p, the tumor suppression effect of cisplatin which is the first-line drug of malignant pleural mesothelioma and the tumor suppression effect of miR-3140-3p were compared in vivo.
Six weeks-old male mice (C-B-17/Icr-scid/scid Jcl) were used as mice. Malignant pleural mesothelioma cell strain EHMES-10 which expresses the luciferase gene was used as the tumor cell.
The protocol is shown below.
The mixed anesthetic drug was prepared as in Table 8 shown above. The miRNA/A6K mixture was prepared as in Tables 9 and 10 shown above. Cisplatin was prepared as in Table 11 below.
Note that the same control RNA as that used in Experiment 7 was employed as the control.
Experimental results are shown in
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20220218733 A1 | Jul 2022 | US |
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Parent | 16760817 | US | |
Child | 17694982 | US |