CN117802096A - Small RNA and small RNA composition of anti-hepatoma bolus for preventing and/or treating hepatoma, and preparation method and application thereof - Google Patents
Small RNA and small RNA composition of anti-hepatoma bolus for preventing and/or treating hepatoma, and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a small RNA and a small RNA composition of a PIANZAIHUANG for preventing and/or treating liver cancer, and a preparation method and application thereof, and belongs to the technical field of biological medicines. The small RNA of the invention is PTH-sRNA-6 with the nucleotide sequence shown as SEQ ID NO.1 and PTH-sRNA-24 with the nucleotide sequence shown as SEQ ID NO. 2. The small RNA composition consists of PTH-sRNA-6 and PTH-sRNA-24. The 2 small RNA molecules can be effectively absorbed by ingestion, exist in vivo stably, and can be combined with and inhibit the expression of NDRG3, CUX1, CAT1 and SLC7A1 gene mRNA, thereby exerting the treatment and/or prevention effect on liver cancer. The small RNA composition obtained by combining PTH-sRNA-6 and PTH-sRNA-24 can also effectively inhibit the proliferation of liver cancer cells. The 2 small RNA molecules can be used as markers to control the quality of the Pianzaihuang. The research of the invention discovers that the small RNA in the Pianzaihuang is beneficial to treating liver cancer, which has important significance for further research and development and quality control of the Pianzaihuang.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to a small RNA and a small RNA composition of a PIANZAIHUANG for preventing and/or treating liver cancer, a preparation method and application thereof.
Background
Liver cancer is one of the most common malignant tumors of the digestive system, the fourth most lethal tumor worldwide, constitutes a serious threat to human health, and has higher morbidity especially in eastern asia, africa and south europe. Due to the characteristics of complex pathogenesis, hidden early course, high morbidity, poor prognosis, high recurrence and high metastasis after operation, and the like, particularly the lack of an effective early diagnosis method and an effective treatment means leads to high patient mortality. At present, early surgical excision is the most main and effective treatment means for liver cancer, but most patients enter middle and late stages when diagnosed for the first time, so that surgical treatment is greatly limited, and the recurrence rate of the surgery is high. In addition, the treatment methods such as local ablation, radiotherapy and chemotherapy, interventional therapy, liver transplantation and the like are limited by a plurality of contraindications in clinical application, and the overall curative effect is still limited.
The pathogenesis of the tumor is considered as dynamic change process of toxin, blood stasis and deficiency in the traditional Chinese medicine, and the poisoning and the blood stasis are dominant in the disease development, so that the method is a key for preventing and treating the tumor. The Pianzaihuang is a lozenge refined by various rare traditional Chinese medicines such as bezoar, pseudo-ginseng, snake gall, musk and the like, and the patent application with publication number of CN104189037A discloses the application of the Pianzaihuang in resisting tumor, which indicates that the Pianzaihuang has a certain therapeutic effect on liver cancer. However, the mechanism of the action of the Pianzaihuang is not clear.
Small RNA (sRNA) refers to a class of non-coding RNA molecules, including micro RNA (miRNA), small interference RNA (siRNA), piwi-interacting RNA (piRNA), typically below 200nt in length. The small RNA can regulate gene expression at the posttranscriptional level and plays an important role in physiological and pathological processes. Recently, research has found that small RNA in part of traditional Chinese medicines can enter blood and tissue organs of a human body through alimentary canal and play an important role in disease treatment. For example, MIR2911 in the honeysuckle decoction can enter the body of a mouse in a gastric feeding way, and can directly act on influenza virus. These findings suggest that the small RNA of the traditional Chinese medicine can be taken orally as a novel traditional Chinese medicine active ingredient which is ignored for a long time and can be absorbed through the alimentary canal to reach the target organ to exert the therapeutic value.
Disclosure of Invention
The invention aims to provide a small RNA and a small RNA composition of a PIANZAIHUANG for preventing and/or treating liver cancer, and a preparation method and application thereof.
The invention provides a small RNA for preventing and/or treating liver cancer, which is a nucleotide sequence shown as SEQ ID NO.1 or SEQ ID NO. 2.
The invention also provides application of the small RNA in preparing a medicament for preventing and/or treating liver cancer.
Further, the medicine is a medicine for inhibiting proliferation of liver cancer cells;
preferably, the medicament prepared by the nucleotide sequence shown in SEQ ID NO.1 is a medicament for inhibiting the expression of a CUX1 gene;
and/or the medicine prepared by the nucleotide sequence shown in SEQ ID NO.2 is a medicine for inhibiting the expression of NDRG3, CAT1 and SLC7A1 genes.
The invention also provides a small RNA composition for preventing and/or treating liver cancer, which is composed of nucleotide sequences shown as SEQ ID NO.1 and SEQ ID NO.2 as components.
Further, the mol percent of the nucleotide sequence shown in SEQ ID NO.1 is 60-80%, and the mol percent of the nucleotide sequence shown in SEQ ID NO.2 is 20-40%.
Further, the mol percent of the nucleotide sequence shown in SEQ ID NO.1 is 76-77%, and the mol percent of the nucleotide sequence shown in SEQ ID NO.2 is 23-24%.
Further, the nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO.2 have a molar ratio of 1:0.3.
The invention also provides a preparation method of the small RNA composition, which comprises the following steps:
mixing the nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO. 2.
The invention also provides application of the small RNA composition in preparing a medicament for preventing and/or treating liver cancer.
Further, the medicine is a medicine for inhibiting proliferation of liver cancer cells; and/or the medicine is a medicine for promoting apoptosis of liver cancer cells;
preferably, the drug is a drug for inhibiting the expression of NDRG3, CUX1, CAT1 and SLC7A1 genes.
The invention also provides a medicine for preventing and/or treating liver cancer, which is a preparation prepared by taking the small RNA or the small RNA composition as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients.
Compared with the prior art, the invention has the beneficial effects that:
the invention discovers 2 kinds of small RNAs (PTH-sRNA-6 and PTH-sRNA-24) from the traditional Chinese patent medicine PIANZAIHUANG, and the 2 kinds of small RNAs have inhibition effect on lung cancer target genes, have the effect of inhibiting liver cancer cell proliferation, and can be used for preventing and/or treating liver cancer. In addition, the composition combined by the 2 small RNAs can also effectively inhibit the growth of liver cancer cells, and has good prevention and/or treatment effects on liver cancer. Meanwhile, 2 kinds of small RNAs can be effectively absorbed through ingestion, exist stably and are convenient and effective to use. The 2 small RNAs can be used as markers to control the quality of the Pianzaihuang, and the Pianzaihuang with high content of 2 small RNAs has better effect of treating liver cancer. The research of the invention discovers that the small RNA in the Pianzaihuang is beneficial to treating liver cancer, which has important significance for further research and development and quality control of the Pianzaihuang.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
FIG. 1 is a flow chart of a small RNA high throughput sequencing experiment.
FIG. 2 shows Solexa sequenced small RNA copy numbers.
FIG. 3 shows the levels of the small Pitaze RNA PTH-sRNA-6 and PTH-sRNA-24 in the serum of each group of mice.
FIG. 4 shows the levels of the small Pitaze further RNA PTH-sRNA-6 and PTH-sRNA-24 in the livers of the mice in each group.
FIG. 5 shows the binding capacity of PTH-sRNA-6 to CUX1 gene mRNA.
FIG. 6 shows the binding capacity of PTH-sRNA-24 to mRNA of NDRG3 gene.
FIG. 7 shows the binding capacity of PTH-sRNA-24 to mRNA of CAT1 gene.
FIG. 8 shows the binding capacity of PTH-sRNA-24 to mRNA of SLC7A1 gene.
Fig. 9 shows the binding ability of small RNA of small bolus to target site through luciferase reporter assay.
FIG. 10 shows the flow cytometry detection of apoptosis rate of liver cancer cells.
FIG. 11 shows the detection of proliferation of hepatoma cells by CCK 8.
Fig. 12 shows tumor diameter change in nude mice.
Detailed Description
The materials and equipment used in the embodiments of the present invention are all known products and are obtained by purchasing commercially available products.
The Tze Huang used in the invention is provided by Zhangzhou Tze Huang pharmaceutical industry Co., ltd.
The inventor firstly extracts the total RNA of the Tze Huang and carries out the high-flux sequencing of the small RNA; and then, carrying out intragastric bolus on the mice, extracting RNA from the serum of the mice, carrying out high-throughput sequencing on the small RNA, screening the small RNA with the medicine source of the bolus, and selecting the small RNA with the copy number larger than 100. Through researches, the 2 small RNAs related to liver cancer are found.
Example 1 verification of 2 Small RNAs of the invention in Pianzaihuang
1. Extraction of total RNA in Pianzaihuang
Taking a proper amount of the Tze Huang, grinding the Tze Huang into 100 meshes of powder at normal temperature or low temperature, total RNA from the samples was extracted with Trizol reagent. The specific extraction method comprises the following steps:
1) Grinding Tze Huang into powder by using a low-temperature grinder, weighing 50mg, putting into a 1.5ml centrifuge tube, adding 1ml of trizol vortex, and standing on ice for 10min;
2) Adding 200 μl of chloroform, shaking vigorously to mix them, and standing for 5min;
3) Centrifuging at 4deg.C and 14000g for 20min;
4) Sucking the supernatant into a new 1.5ml centrifuge tube, adding isopropanol with the volume of 1-2 times of the supernatant, uniformly mixing, and then placing into a-20 ℃ for precipitation for more than 1 hour;
5) Centrifuging at 4deg.C and 14000g for 20min;
6) Discarding the supernatant precipitate, adding 1ml of 75% alcohol prepared with DEPC water, and blowing;
7) Centrifuging at 4deg.C and 14000g for 20min;
8) Discarding the supernatant, inverting a 1.5ml centrifuge tube, and airing the alcohol for about 5-10min;
9) Dissolving in DEPC water, and preserving at-80deg.C to obtain total RNA of PIANZAIHUANG.
2. Detection of the content of 2 kinds of small RNAs of the invention
After total RNA extraction of the Tze Huang, detection can be performed by high throughput sequencing techniques, reverse transcription PCR (RT-PCR), real-time fluorescent quantitative PCR (qPCR), RNA chips, northern Blotting, in situ hybridization, and the like. The specific method of the embodiment is as follows:
extracting total RNA from Tze Huang, performing PAGE electrophoresis to recover 18-45nt RNA molecule, adding 3' joint, adding RT primer with UMI into the system, and performing reverse transcription extension to synthesize cDNA chain. The following 2 small RNA sequences (Table 1) and the content thereof in the Pitahuang were detected by conventional small RNA high-throughput sequencing (experimental flow is shown in FIG. 1). The RT primer with UMI is supplied by sequencing company (Shenzhen megagene Co., ltd.).
TABLE 1 names and sequences of small RNAs
The 2 small RNA copy numbers are shown in FIG. 2. As can be seen from fig. 2 and table 1: the content of 2 small RNAs in the Tze Huang is higher. The content of PTH-sRNA-6 in the Pitahuang is 256786 copies, and the content of PTH-sRNA-24 in the Pitahuang is 77153 copies.
The beneficial effects of the present invention are demonstrated by specific test examples below.
Test example 1 stability study of the small RNA of Pianzaihuang of the invention into the body by ingestion
1. Experimental method
And detecting small RNA of the Pianzaihuang which enters the animal body through ingestion and exists stably by using an RT-qPCR method. Male C57 mice were randomly divided into PBS control group, single administration group of PIANZAIHUANG, and multiple administration group of PIANZAIHUANG. After 12h of fasted food, the Tze Huang is ground into 100-mesh powder, and then is prepared into suspension by PBS for gastric administration. The single administration group is only administrated by intragastric administration once, and the multiple administration group is administrated by intragastric administration once every 3 hours for three times. The dosage of the bolus for each time is 0.1g/100g. The PBS control group was perfused with an equivalent dose of PBS. After the last gastric lavage is finished for 6 hours, blood of the mice is taken to separate serum, and liver tissues are taken. RNA was extracted from serum and liver tissues, and the small RNA content of Pitaze Huang (PTH-sRNA-6 and PTH-sRNA-24 content, respectively) was detected by Real-time PCR. Primers for reverse transcription PCR and qPCR detection of small RNA of Pitahuang are shown in Table 2.
TABLE 2 reverse transcription PCR and qPCR detection primer list
Reverse transcription (10 μl) of sRNA of Pitahuang was performed using the system shown in Table 3.
TABLE 3 sRNA reverse transcription reaction System
After the system is prepared, the mixture is evenly mixed and put into a PCR instrument for reaction, and the reaction procedure is shown in Table 4.
TABLE 4 reverse transcription procedure for sRNA
After the reverse transcription is completed, quantitative detection is performed by using qPCR primers. qPCR system is shown in Table 5 and reaction procedure is shown in Table 6.
TABLE 5 sRNA fluorescent quantitative PCR reaction System
TABLE 6 fluorescent quantitative PCR reaction procedure
2. Experimental results
The relative concentration results of the two small RNAs in the blood serum and liver tissues are shown in fig. 3 and 4: the results show that all 2 small RNAs of the Pianzaihuang can be absorbed through the alimentary canal and reach the serum and liver tissues. Experimental results show that the sRNA of the 2-type Tze Huang can enter animals through ingestion and exist stably.
Test example 2 bioinformatics means analysis of the binding of small RNA of Pianzaihuang to target Gene
1. Experimental method
Whether the small RNA of the Pianzaihuang can be combined with a series of target genes related to liver cancer diseases or not is studied, and the therapeutic value of the small RNA on the liver cancer diseases is exerted by inhibiting the expression of target genes. And predicting the inhibition effect of the 2 small RNAs of the Tze Huang on NDRG3, CUX1, CAT1 and SLC7A1 target genes by using RNA hybrid.
2. Experimental results
The results were as follows:
the binding capacity of PTH-sRNA-6 to the mRNA of the CUX1 gene is shown in FIG. 5, which illustrates the binding pattern, binding site and free energy of binding of PTH-sRNA-6 to the mRNA of the CUX1 gene. Among them, PTH-sRNA-6 has 1 binding mode with CUX 1.
The binding capacity of PTH-sRNA-24 and mRNA of NDRG3, CAT1 and SLC7A1 genes is shown in figures 6-8, and the binding mode, binding site and free binding energy of PTH-sRNA-24 and mRNA of NDRG3, CAT1 and SLC7A1 genes are illustrated. Wherein, there are 3 binding modes of PTH-sRNA-24 and NDRG3, 1 binding mode of PTH-sRNA-24 and CAT1, and 1 binding mode of PTH-sRNA-24 and SLC7A 1.
The four genes NDRG3, CUX1, CAT1 and SLC7A1 are all accepted to play a role in promoting the occurrence and development of liver cancer in the process of treating the liver cancer, and proliferation of liver cancer cells can be inhibited by inhibiting expression of the four target genes, so that the four genes NDRG3, CUX1, CAT1 and SLC7A1 can play a role in treating the liver cancer.
The experimental results show that: PTH-sRNA-6 has an inhibiting effect on CUX1 genes, PTH-sRNA-24 has an inhibiting effect on NDRG3, CAT1 and SLC7A1 genes, which indicates that small RNAs PTH-sRNA-6 and PTH-sRNA-24 have an inhibiting effect on liver cancer cell proliferation, and can be used for preventing and/or treating liver cancer; and the combination of the PTH-sRNA-6 and the PTH-sRNA-24 can simultaneously inhibit the expression of four genes of NDRG3, CUX1, CAT1 and SLC7A1, thereby further playing a role in preventing and/or treating liver cancer.
Test example 3 inhibition effect of small RNA of Pianzaihuang on target Gene
1. Experimental method
Small RNA of two kinds of Twoliving Twostyle bolus including PTH-sRNA-6 and PTH-sRNA-24 obtained from the Tze Huang by the method described in example 1, or can be synthesized directly by conventional techniques.
The inhibition effect of the small RNA of the Pianzaihuang on each target gene is proved by a luciferase report experiment:
the segment sequence fragments (Table 7) of NDRG3, CUX1, CAT1 and SLC7A1 target genes combined with two small RNAs of PTH-sRNA-6 and PTH-sRNA-24 were inserted into pMIR-REPORT Luciferase plasmid respectively to construct luciferase reporter plasmids of the genes. Positive clones were screened and sequenced for verification, clones were amplified and plasmids were purified. The relevant cells are cultured and inoculated in a 24-well plate, and after 12-16 hours, the luciferase expression plasmid carrying the predicted target gene to be detected and small RNA of Pitazaihuang (two sRNAs of PTH-sRNA-6 and PTH-sRNA-24) are co-transfected into the cells. After 24 hours, the cells were collected and lysed, and luciferase substrate was added and luciferase reacted with the substrate to produce luciferin. By detecting the intensity of fluorescence, the luciferase activity can be determined. And comparing with the transfected nonsense small RNA group, so as to judge whether the predicted target point can be inhibited by the small RNA of the Tze Huang. Nonsense small RNAs are nonsensical small RNA sequences that do not bind to the target gene.
Nonsense small RNA sequence: GGCAGCUAACCUAUAUGACAUGC (SEQ ID NO. 9) Table 7. Fragment of the sRNA and target Gene binding sequence of Pianzaihuang
2. Experimental results
The report experimental result of luciferase shows (figure 9) that the small RNA of the anti-inflammatory agent has inhibition effect on four target genes of NDRG3, CUX1, CAT1 and SLC7A 1. The nonsense small RNA has no inhibiting effect on the four target genes. The small RNA composition of the Pianzaihuang has an inhibiting effect on liver cancer target genes, and can be used for preventing and/or treating liver cancer.
Test example 4 apoptosis experiment
1. Experimental method
Two kinds of patches, PTH-sRNA-6 and PTH-sRNA-24 the small RNA of the Huang is directly synthesized by adopting a conventional technology.
The apoptosis experiment proves that the sRNA of the Pianzaihuang can promote the apoptosis of tumor cells: hepG2 cells are paved on a 6-hole plate, PTH-sRNA-6 and PTH-sRNA-24 are co-transfected when the cell density reaches 70% -80%, and apoptosis influence of the sRNA of the Pianzaihuang on liver cancer cells is verified by flow cytometry after 24 hours. The molar ratio of PTH-sRNA-6 to PTH-sRNA-24 was 1:0.3, i.e., 77 mole percent PTH-sRNA-6 and 23 mole percent PTH-sRNA-24 in the mixed small RNA. Normal cells not transfected with sRNA served as controls; the scramble RNA is transfected nonsense small RNA, and the nucleotide sequence of the nonsense small RNA is shown as SEQ ID NO.9 and is used as a negative control.
2. Experimental results
The results of the apoptosis experiments are shown in fig. 10, and the results show that: the apoptosis rate of liver cancer cells is increased after the liver cancer cells are treated by the Tsaoko sRNA composition (PTH sRNA: PTH-sRNA-6 and PTH-sRNA-24). The sRNA composition of the Pianzaihuang can promote apoptosis of liver cancer cells.
Test example 5 cell proliferation assay
1. Experimental method
Two kinds of patches, PTH-sRNA-6 and PTH-sRNA-24 the small RNA of the Huang is directly synthesized by adopting a conventional technology.
Cell proliferation experiments prove that the sRNA of the Pianzaihuang can inhibit proliferation of liver cancer cells: hepG2 cells were plated on 6-well plates, PTH-sRNA-6 and PTH-sRNA-24 were co-transfected at cell densities of 70% -80%, and sRNA-transfected cells (PTH sRNA) and untransfected cells (control) were plated on 96-well plates after 6 hours, and cell proliferation (CCK 8) was examined at 12 hours, 24 hours, 36 hours, 48 hours and 72 hours, respectively. The molar ratio of PTH-sRNA-6 to PTH-sRNA-24 was 1:0.3, i.e., 77 mole percent PTH-sRNA-6 and 23 mole percent PTH-sRNA-24 in the mixed small RNA. Normal cells not transfected with sRNA served as controls; the scramble RNA is transfected nonsense small RNA, and the nucleotide sequence of the scramble RNA is shown as SEQ ID NO.9 and is used as a negative control.
2. Experimental results
The results of the cell proliferation assay are shown in fig. 11, which shows that: the treatment of the Pianzaihuang sRNA composition (PTH sRNA: PTH-sRNA-6 and PTH-sRNA-24) can inhibit proliferation of liver cancer cells. The sRNA composition of the invention has therapeutic effect on liver cancer.
Test example 6 in vivo verification of inhibition of sRNA of Pianzaihuang on liver cancer cell proliferation
1. Experimental method
Transfection of sRNA of Pianzaihuang into liver cancer cells (the molar ratio of PTH-sRNA-6 to PTH-sRNA-24 is 1:0.3, i.e. 77 mol% PTH-sRNA-6 and 23 mol% PTH-sRNA-24 in the mixed small RNA), followed by 1X 10) 6 The nude mice were subcutaneously tumor implanted at individual cell/dose. Tumor diameter changes were measured with vernier calipers on days 3, 6, 9, 12, 15, 18 after tumor implantation. The liver cancer cells not transfected with the Pianzaihuang sRNA are used for subcutaneous tumor implantation of nude mice as a control group. The scramble RNA is transfected nonsense small RNA, and the nucleotide sequence of the scramble RNA is shown as SEQ ID NO.9 and is used as a negative control.
2. Experimental results
The results of the tumor diameter change of the nude mice are shown in FIG. 12, and the results show that: compared with the untreated hepatoma cell tumor implantation result (control), the proliferation rate of cells treated with the sRNA compositions of Tback (PTH-sRNA-6 and PTH-sRNA-24) was slowed, and the volume increase was slower. The invention shows that the small RNA composition of the PIANZAIHUANG can effectively inhibit the growth of liver cancer tumors.
In conclusion, 2 kinds of small RNAs (PTH-sRNA-6 and PTH-sRNA-24) are found from the traditional Chinese patent medicine PIANZAIHUANG, and the 2 kinds of small RNAs have an inhibitory effect on lung cancer target genes, have the effect of inhibiting proliferation of liver cancer cells, and can be used for preventing and/or treating liver cancer. In addition, the composition combined by the 2 small RNAs can also effectively inhibit the growth of liver cancer cells, and has good prevention and/or treatment effects on liver cancer. Meanwhile, 2 kinds of small RNAs can be effectively absorbed through ingestion, exist stably and are convenient and effective to use. The 2 small RNAs can be used as markers to control the quality of the Pianzaihuang, and the Pianzaihuang with high content of 2 small RNAs has better effect of treating liver cancer. The research of the invention discovers that the small RNA in the Pianzaihuang is beneficial to treating liver cancer, which has important significance for further research and development and quality control of the Pianzaihuang.
Claims (11)
1. A small RNA for preventing and/or treating liver cancer, characterized in that: it is a nucleotide sequence shown as SEQ ID NO.1 or SEQ ID NO. 2.
2. Use of the small RNA of claim 1 for the preparation of a medicament for the prevention and/or treatment of liver cancer.
3. Use according to claim 2, characterized in that: the medicine is used for inhibiting proliferation of liver cancer cells;
preferably, the medicament prepared by the nucleotide sequence shown in SEQ ID NO.1 is a medicament for inhibiting the expression of a CUX1 gene;
and/or the medicine prepared by the nucleotide sequence shown in SEQ ID NO.2 is a medicine for inhibiting the expression of NDRG3, CAT1 and SLC7A1 genes.
4. A small RNA composition for preventing and/or treating liver cancer, characterized in that: it is composed of the nucleotide sequences shown as SEQ ID NO.1 and SEQ ID NO.2 as components.
5. The small RNA composition of claim 4, wherein: the mol percent of the nucleotide sequence shown in SEQ ID NO.1 is 60-80%, and the mol percent of the nucleotide sequence shown in SEQ ID NO.2 is 20-40%.
6. The small RNA composition of claim 5, wherein: the mol percent of the nucleotide sequence shown in SEQ ID NO.1 is 76-77%, and the mol percent of the nucleotide sequence shown in SEQ ID NO.2 is 23-24%.
7. The small RNA composition of any one of claims 4 to 6, wherein: the mol ratio of the nucleotide sequences shown in SEQ ID NO.1 to SEQ ID NO.2 is 1:0.3.
8. The method for producing a small RNA composition of any one of claims 4 to 7, wherein: it comprises the following steps:
mixing the nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO. 2.
9. Use of the small RNA composition of any one of claims 4 to 7 for the preparation of a medicament for the prevention and/or treatment of liver cancer.
10. Use according to claim 9, characterized in that: the medicine is used for inhibiting proliferation of liver cancer cells; and/or the medicine is a medicine for promoting apoptosis of liver cancer cells;
preferably, the drug is a drug for inhibiting the expression of NDRG3, CUX1, CAT1 and SLC7A1 genes.
11. A medicament for preventing and/or treating liver cancer, which is characterized in that: the preparation is prepared by taking the small RNA as claimed in claim 1 or the small RNA composition as claimed in any one of claims 4 to 7 as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients.
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